See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/318921610

PROTEIN AND POLYPEPTIDE HORMONES (Medicinal Chemistry) by: Dr Asif

Husain

Chapter · August 2017

CITATIONS READS

0 3,452

1 author:

Asif Husain
Jamia Hamdard University
286 PUBLICATIONS   3,395 CITATIONS   

SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Furanones and pyrrolones View project

Phytochemistry and potential therapeutic actions of Boswellic acid View project

All content following this page was uploaded by Asif Husain on 05 August 2017.

The user has requested enhancement of the downloaded file.

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

PROTEIN AND POLYPEPTIDE HORMONES

(Medicinal Chemistry Theory)

For
B. Pharmacy students

BY
DR. ASIF HUSAIN
Associate Professor
DEPARTMENT OF PHARM. CHEMISTRY
SCHOOL OF PHARMACEUTICAL EDUCATION AND
RESEARCH (SPER)
JAMIA HAMDARD

Protein and Polypeptide Hormones 1

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

e-Book title: Medicinal Chemistry

Chapter title: Protein and Polypeptide Hormones

Author name: Dr. Asif Husain

Suggested reading: 1. K.D. Tripathi: Essentials of Medical Pharmacology.

2. Goodman Gilman`s: The Pharmacological basis of Therapeutics by
Alfred Goodman Gilman.
3. R.F. Doerge: Wilson & Gisvold`s Text Book of Organic and
Pharmaceutical Chemistry, J. Lippincott Co., Philadelphia.
4. H.P. Rang and M.M. Dale: Pharmacology
5. W.O. Foye: Principles of Medicinal Chemistry, Lea & Febiger,
Philadelphia.
6. www.pubmed.com
6. www.google.com

Keywords: Corticotropic hormone, somatotropin, hCG, FSH, LH, TSH, insulin,

glucagon, GIT-hormones, oxytocin, vasopressin, PTH, peptides.

Protein and Polypeptide Hormones 2

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

Protein and Polypeptide Hormones

NOMENCLATURE

1. General Principles

Naturally occurring oligo- and polypeptides are generally referred to by trivial names; their

systematic names are so cumbersome that they are of little use. Most of the peptide hormones

already have well-established trivial names indicating either natural source (e.g. insulin) or

physiological action (e.g. relaxin, prolactin). However, some of the trivial names are so long that

these hormones are known mainly by abbreviations (e.g. FSH for follicle-stimulating hormone).

This is unfortunate, and it was therefore considered advisable to create suitable names for those

peptide hormones not already having well established short trivial names. Three principles have

been observed:

(a) New names for hormones of the adenohypophysis bear the ending "-tropin";

(b) Hypothalamic releasing factors (hormones) bear the ending "-liberin";

(c) Hypothalamic release-inhibiting factors (hormones) bear the ending "-statin".

2. Trivial Names

The trivial names proposed for peptide hormones are given in the "Appendix." Abbreviations of

the new names are not proposed, and the use of currently fashionable abbreviations is

discouraged.

3. Species Designation

Since peptide hormones show species variation in their amino-acid sequence, their names are

essentially "generic names", and are insufficient to specify a single chemical compound. It is

Protein and Polypeptide Hormones 3

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

therefore recommended that authors add to the name of each hormone the species from which

the hormone was isolated, or at least indicate the biological source(s) where appropriate.

4. Special Groups of Hormones

a) Hypothalamic Factors (Hormones) - The hypothalamic "releasing factors" or "releasing

hormones" have no well established trivial names. It is recommended that the trivial names given

in the "Appendix" be used for the releasing factors (hormones). They are based on the ending "-

liberin" added to the prefix of the pituitary hormone released by the factor. Thus, "thyroliberin"

indicates the hypothalamic peptide stimulating the release (and perhaps also the biosynthesis) of

thyrotropin, the corresponding tropic hormone, from the pituitary gland. (Note that the ending

"-tropin" is no longer retained in the name; it is implied in the definition of "-liberin").

The names of those factors inhibiting the release (and perhaps the synthesis) of pituitary

hormones are formed in a similar way with the suffix "-statin".

(b) Pituitary Hormones - Most of the hormones of the adenohypophysis have acceptable trivial

names ending in -tropin. Follicle-stimulating hormone is known as "follitropin," and luteinizing

hormone as "lutropin." It is recommended that pituitary hormones discovered in the future also

be named with the ending -tropin, This suffix is restricted to pituitary and similar hormones and

should not be used for, e.g. crustacean hormones acting on pigment cells.

Some placental hormones are physiologically very similar to pituitary hormones. They are

named accordingly with the prefix "chorio-", e.g. choriogonadotropin for chorionic

gonadotropin.

Protein and Polypeptide Hormones 4

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

(c) Invertebrate Peptide Hormones -Though some of the invertebrate peptide hormones have

been isolated in pure form and their amino-acid compositions have been determined, the field

has not yet developed to a stage where a list of names seems warranted.

It is, however, recommended that the suffixes defined above for hypothalamic and pituitary

hormones are not used in a different sense in invertebrates. Thus, a crustacean color change

hormone acting on, e.g. erythrophores, should not be named "erythrotropin," a hormone causing

release of eggs and/or sperm in sea urchins should not be called "gametoliberin."

Appendix. List of Peptide Hormonesa

Current
Trivial name Other names
abbreviationb
1. Hypothalamic Factors

Corticoliberin Corticotropin-releasing factor CRF

Folliberin Follicle-stimulating-hormone-releasing factor FSH-RF
Gonadoliberinc Gonadotropin-releasing factor (LH/FSH-RF)
Luliberin Lutemizing hormone-releasing factor LH-RF (LRF)
Melanoliberin Melanotropin-releasing factor MFR
Melanostatin Melanotropin release-inhibiting factor MIF
Prolactoliberin Prolactin-releasing factor PRF
Prolactostatin Prolactin release-inhibiting factor PIF
Somatotropin-releasing factor;
SRF
Somatoliberin
growth hormone-releasing factor GH-RF

Somatostatin Somatotropin release-inhibiting factor

Thyroliberin Thyrotropin-releasing factor TRF

2. Pituitary and Related Hormones

Choriogonadotropind Chorionic gonadotropin CG

Protein and Polypeptide Hormones 5

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

Choriomammotropin Chorionic somatomammotropin CS

Corticotropin Adrenocorticotropic hormone

Follitropin Follicle-stimulating hormone FSH

Gonadotropine Gonadotropin hormone

Glumitocinf Ocytocinj

Isotocing Ocytocinj

Lipotropin Lipotropic hormone LPH

Lutropin Luteinizing hormone;
LH
(Interstitial cell-stimulating hormone) (ICSH)

Melanotropinh Melanocyte-stimulating hormone MSH

j
Mesotocini Ocytocin

Ocytocinj (Oxytocin) OXT

Mammatropic hormone; mammatropin;
Prolactin PRL
lactotropic hormone; lactotropin
Somatropic hormone;
STH
Somatotropin
growth hormone GH

Thyrotropin Thyrotropic hormone TSH

Urogonadotropink (Human) Menopausal gonadotropin HMG
Vasopressin Adiuretin; antidiuretic hormone VP, ADH
Vasotocin Ocytocinj

3. Other Peptide Hormones

Angiotensin Angiotensin II

Bradykinin Kinin-9

Calcitonin Thyrocalcitonin

Erythropoietin
Gastrin
Gastrin sulphate Gastrin II

Glucagon Hyperglycemic factor (HGF)

Insulin

Protein and Polypeptide Hormones 6

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

Kallidin Kinin-10

Pancreozymin Cholecystokinin

Parathyrinl Parathyroid hormone; Parathormone

Proangiotensin Angiotensin I

Relaxin
Secretin
Somatomedinm Sulfation factor

Thymopoietinn Thymin

a
For convenience, some biologically active peptides that may not fulfill all criteria of a hormone

are included.
b
Abbreviations, old or new, are not recommended; they are given here for identification
purposes only.
c
This name indicates a hypothalamic substance releasing gonadotropin. It may also be used for

the decapeptide isolated from pig hypothalami and known as luteinizing hormone/follicle-

stimulatinghormone releasing factor, abbreviated LH/FSH-RF,b since the peptide induces the

release of both lutropin and follitropin in constant proportions and thus carries the activity of

both luliberin and folliberin (see also Footnote e).

d
The chorionic gonadotropins have in most species (including man) the action of both follitropin

and lutropin and are therefore termed "gonadotropins."

e
Gonadotropin is to be used for hormones having the activity of both follitropin and lutropin,

like the gonadotropins of cold-blooded vertebrates. It may also be used for impure preparations

containing lutropin and follitropin.

f
In elasmobranch fishes.
g
In bony fishes.

h
Two peptides have been sequenced and designated -melanotropin and -melanotropin.

Protein and Polypeptide Hormones 7

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

i
In birds and reptiles.

j
The name of this hormone is derived from Greek  (OKYTOKOS = fast birth, prompt

delivery), not from the Greek  (oxys = acid; fast). The spelling ocytocin should therefore be

preferred; moreover, it avoids confusion with oxy, meaning "related to oxygen." However,

oxytocin is in wide use, especially in the English language. Therefore, both spellings are listed as

optional.

k
Most work has been done on the human hormone, known as Human Menopausal Gonadotropin

(HMG); it is a pitutary hormone, chemically changed during passage through the kidney. Due to

its occurrence in urine, it has been termed "urogonadotropin."

l
Parathyrin is a new name suggested here. The synonym Parathormone is a proprietary name.
m
The name "somatomedin" was suggested by a group working in the field (Nature 235, 107

(1972)).
n
A polypeptide from the thymus. The name proposed was suggested in a letter to Nature (249,

863 (1974) to avoid confusion with the earlier "thymine" from nucleic acids. "Thymin" should be

abandoned.

Structure and Function of Hormones

The integration of body functions in humans and other higher organisms is carried out by the

nervous system, the immune system, and the endocrine system. The endocrine system is

composed of a number of tissues that secrete their products, called endocrine hormones, into the

circulatory system; from there they are disseminated throughout the body, regulating the function

of distant tissues and maintaining homeostasis. In a separate but related system, exocrine tissues

secrete their products into ducts and then to the outside of the body or to the intestinal tract.

Protein and Polypeptide Hormones 8

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

Classically, endocrine hormones are considered to be derived from amino acids, peptides, or

sterols and to act at sites distant from their tissue of origin. However, the latter definition has

begun to blur as it is found that some secreted substances act at a distance (classical endocrines),

close to the cells that secrete them (paracrines), or directly on the cell that secreted them

(autocrines). Insulin-like growth factor-I (IGF-I), which behaves as an endocrine, paracrine,

and autocrine, provides a prime example of this difficulty.

Hormones are normally present in the plasma and interstitial tissue at concentrations in the range

of 10-7M to 10-10M. Because of these very low physiological concentrations, sensitive protein

receptors have evolved in target tissues to sense the presence of very weak signals. In addition,

systemic feedback mechanisms have evolved to regulate the production of endocrine hormones.

Once a hormone is secreted by an endocrine tissue, it generally binds to a specific plasma protein

carrier, with the complex being disseminated to distant tissues. Plasma carrier proteins exist for

all classes of endocrine hormones. Tissues capable of responding to endocrines have 2 properties

in common: they posses a receptor having very high affinity for hormone, and the receptor is

coupled to a process that regulates metabolism of the target cells. Receptors for most amino acid-

-derived hormones and all peptide hormones are located on the plasma membrane. Activation of

these receptors by hormones (the first messenger) leads to the intracellular production of a

second messenger, such as cAMP, which is responsible for initiating the intracellular biological

response. Steroid and thyroid hormones are hydrophobic and diffuse from their binding proteins

in the plasma, across the plasma membrane to intracellularly localized receptors. The resultant

Protein and Polypeptide Hormones 9

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

complex of steroid and receptor bind to response elements of nuclear DNA, regulating the

production of mRNA for specific proteins.

Receptors for Peptide Hormones

With the exception of the thyroid hormone receptor, the receptors for amino acid-derived and

peptide hormones are located in the plasma membrane. Receptor structure is varied: some

receptors consist of a single polypeptide chain with a domain on either side of the membrane,

connected by a membrane-spanning domain. Some receptors are comprised of a single

polypeptide chain that is passed back and forth in serpentine fashion across the membrane,

giving multiple intracellular, transmembrane, and extracellular domains. Other receptors are

composed of multiple polypeptides. For example, the insulin receptor is a disulfide-linked

tetramer with the β subunits spanning the membrane and the α subunits located on the exterior

surface.

Subsequent to hormone binding, a signal is transduced to the interior of the cell, where second

messengers and phosphorylated proteins generate appropriate metabolic responses. The main

second messengers are cAMP, Ca2+, inositol triphosphate (IP3) , and diacylglycerol (DAG) .

Proteins are phosphorylated on serine and threonine by cAMP-dependent protein kinase (PKA)

and DAG-activated protein kinase C (PKC). Additionally a series of membrane-associated and

intracellular tyrosine kinases phosphorylate specific tyrosine residues on target enzymes and

other regulatory proteins.

Protein and Polypeptide Hormones 10

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

The hormone-binding signal of most, but not all, plasma membrane receptors is transduced to the

interior of cells by the binding of receptor-ligand complexes to a series of membrane-localized

GDP/GTP binding proteins known as G-proteins. When G-proteins bind to receptors, GTP

exchanges with GDP bound to the α subunit of the G-protein. The Gα -GTP complex binds

adenylate cyclase, activating the enzyme. The activation of adenylate cyclase leads to cAMP

production in the cytosol and to the activation of PKA, followed by regulatory phosphorylation

of numerous enzymes. Stimulatory G-proteins are designated Gs, inhibitory G-proteins are

designated Gi.

A second class of peptide hormones induces the transduction of 2 second messengers, DAG and

IP3. Hormone binding is followed by interaction with a stimulatory G-protein, which is followed

in turn by G-protein activation of membrane-localized phospholipase C-γ, (PLC- γ). PLC- γ

hydrolyzes phosphatidylinositol 4,5-bisphosphate (PIP2) to produce 2 messengers: IP3, which is

soluble in the cytosol, and DAG, which remains in the membrane phase. Cytosolic IP 3 binds to

sites on the endoplasmic reticulum, opening Ca2+ channels and allowing stored Ca2+ to flood the

cytosol. There it activates numerous enzymes, many by activating their calmodulin or

calmodulin-like subunits. DAG has 2 roles: it binds and activates protein kinase C (PKC), and it

opens Ca2+ channels in the plasma membrane, reinforcing the effect of IP 3. Like PKA, PKC

phosphorylates serine and threonine residues of many proteins, thus modulating their catalytic

activity.

Hormones of Hypothalamic Origin

The hypothalamus, which is a relatively small organ that is located in the brain and responsible

for thermoregulation, among other functions, is the secretory source of a number of peptide

Protein and Polypeptide Hormones 11

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

hormones that are transported to the pituitary gland situated immediately below it. These

hormones regulate the synthesis of other peptide hormones produced by the anterior pituitary

(adenohypophysis), and are thus called releasing hormones (RH) or releasing factors (RF), or

inhibitory factors (IF), as the case may be. The release of these hypothalamic hormones is

regulated via cholinergic and dopaminergic stimuli from higher brain centres, and their synthesis

and release controlled by feedback mechanisma from their target organs.

Thyroliberin (Thyrotropin-Releasing hormone; TRH) is the hypothalamic hormone responsible

for the release of the pituitary’s thyrotropin. Thyrotropin stimulates thyroxine and liothyroninr

by the thyroid. The latter thyroid hormones, by feedback regulation, inhibit the action of TRH on

pituitary. Thyroliberin is relatively simple tripeptide that has been characterized as pyroglutamyl-

histidyl-prolinamide. TRH possesses interesting biological properties. in addition to stimulating

the release of thyrotropin, it promotes the release of prolactin. It also has some central nervous

system effects that have been evaluated antidepressant therapeutic potential.

Gonadoliberin, as the name implies, is the gonadotropin-releasing hormone (Gn-RH) (Fig.1),

also known as luteinizing hormone-rleasing hormone (LH-RH). This hypothalamic decapeptide

stimulates the the releasing of luteinizing hormone (LH) and follicle-stimulaing hormone (FSH)

by the pituitary. LH-RH is considered to be of potential therapeutic importance in the treatment

of hypogonadotropic infertility in both males and females.

1 5 6 9 10
pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2

Fig. 1

Protein and Polypeptide Hormones 12

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

It is known that GnRH can be degraded by preferential enzymatic cleavage between Tyr 5-Gly6

and Pro9-Gly10. SAR studies of GnRH have shown that when Gly6 is replaced with certain amino

acids, as well as with changes in the peptide C-terminus, they usually undergo a reduced attack

by proteolytic enzymes, resulting in a longer-lasting action and, for that reason, are referred to as

superagonists. Moreover, when these D-amino acids at position 6 are hydrophobic, the half-life

is enhanced.

Somatostatin is a tetradecapeptide possessing a disulfide bond linking two cysteine residues, 3-

14, in the form of a 38-member ring (Fig. 2). Somatostatin suppresses several endocrine systems.

It inhibits the release of somatotropin and thyrotropin by the pituitary. It also inhibits the

secretion of insulin and glucagons by the pancreas. Gastrin, pepsin and secretin are intestinal

hormones that are likewise affected by somatostatin.

S S
3 14
Ala-Gly-Cys-Lys-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-Cys

Fig. 2

Somatostatin has a shorter half-life (less than 3 minutes), and this, unfortunately, restricts its use

as a therapeutic agent. Many derivatives of somatostatin have been prepared in order to increase

its duration of action or to augment its selectivity of action. The culmination of these SAR

studies has led to the development of octreotide acetate

(Fig. 3), a longer-acting octapeptide analog of somatoatatin, having a half-life of about 1.5 hours.

S S

D-Phe-Cys-Phe-D-Trp-Lys-Thr-Cys-Thr-ol
Fig. 3

Protein and Polypeptide Hormones 13

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

Growth Hormone Releasing Factor (GRF) is a 44-residue-containing peptide, found in minute

quantities in the hypothalamus. It is a positive effector in that it stimulates pituitary release of

somatotropin.

Other hypothalamic hormones include the luteinizing hormone release-inhibiting factor

(LHRIF), prolactin-releasing factor (PRF), corticotropin-releasing factor (CRF),

melanocyte-stimulating hormone –releasing factor (MRF), and melanocyte-stimulating

hormone release-inhibiting factor (MIF).

Pituitary Hormones

The pituitary gland plays a major role in regulating activity of the endocrine organs, including

the adrenal cortex, the gonads, and the thyroid. The posterior pituitary is responsible for the

storage and secretion of hormones vasopressin and oxytocin, controlled by nerve impulses

traveling from the hypothalamus. The anterior pituitary is under the control of hypothalamic

regulatory hormones, and it secretes adrenocorticotropic hormone (ACTH), growth hormone

(GH), LH, FSH, prolactin, and others.

Table-1 summarizes the major hormones synthesized and secreted by the pituitary gland, along

with summary statements about their major target organs and physiologic effects.

Protein and Polypeptide Hormones 14

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

Table-1: Major hormones synthesized and secreted by the pituitary gland.

Major
Hormone target Major Physiologic Effects
organ(s)
Liver, Promotes growth (indirectly), control
Growth hormone adipose of protein, lipid and carbohydrate
tissue metabolism

Thyroid-stimulating Thyroid Stimulates secretion of thyroid

hormone gland hormones

Adrenal
Adrenocorticotropic Stimulates secretion of
gland
Anterior hormone glucocorticoids
(cortex)
Pituitary
Mammary
Prolactin Milk production
gland

Luteinizing Ovary and

Control of reproductive function
hormone testis

Follicle-stimulating Ovary and

Control of reproductive function
hormone testis
Antidiuretic
Kidney Conservation of body water
hormone
Posterior
Pituitary Ovary and Stimulates milk ejection and uterine
Oxytocin
testis contractions

The cells that secrete thyroid-stimulating hormone do not also secrete growth hormone, and they

have receptors for thyroid-releasing hormone, not growth hormone-releasing hormone.

Careful examination of the pituitary gland reveals that it composed of two distinctive parts:

Protein and Polypeptide Hormones 15

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

 The anterior pituitary or adenohypophysis is a classical gland composed predominantly

of cells that secrete protein hormones.

 The posterior pituitary or neurohypophysis is not really an organ, but an extension of the

hypothalamus. It is composed largely of the axons of hypothalamic neurons, which extend

downward as a large bundle behind the anterior pituitary. It also forms the so-called

pituitary stalk, which appears to suspend the anterior gland from the hypothalamus.

Adrenocorticotropic Hormone (ACTH, corticotrophin)

ACTH is a very extensively studied single-chain peptide of 39 residues (Fig. 4). ACTH is

secreted from the anterior pituitary in response to corticotropin-releasing hormone (CRH) from

the hypothalamus, and is derived from a much larger precursor protein known as pro-

opiomelanocortin, the later is the precursor of the melanocyte-stimulating hormones (MSH). As

the name implies, its major action is to regulate the function of the adrenal cortex. More

specifically, it stimulates secretion of glucocorticoids such as cortisol, and has little control over

secretion of aldosterone, the other major steroid hormone from the adrenal cortex.

1 5 10
Ser-Tyr-Ser-Met-Glu-His-Phe-Arg-Tyr-Gly
20 15
Val-Pro-Arg-Arg-Lys-Lys-Gly-Val-Pro-Lys
25 30
Lys-Val-Tyr-Pro-Asn-Gly-Ala-Glu-Asp-Gln
35
Phe-Glu-Leu-Pro-Phe-Ala-Glu-Ala-Ser

Fig. 4

Protein and Polypeptide Hormones 16

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

The SAR studies of ACTH revealed that the COOH-terminal sequence is not particularly

significant for biological activity. Removal of NH2-terminal amino acid results in complete loss

of steroidogenic activity. Complete activity has been reported for synthetic peptides containing

the first 20 amino acids. A peptide containing 24 amino acids has full steroidogenic activity,

without allergenic reactions. This is significant since natural ACTH preparations at times cause

clinically dangerous allergic reactions.

ACTH has a direct anti-inflammatory effect. It is not as potent as other anti-inflammatory agents,

but has fewer side effects when the drug is withdrawn. ACTH may be better for long-term, low

dose therapy in diseases such as rhematoid arthritis, but it is normally prescribed for the

treatment of multiple sclerosis. Withdrawal effects do exists, and they include malaise and

intracranial hypertension.

Melanocyte stimulating hormone (MSH, Melanotropins) has no known function in mammals,

but in reptiles and amphibians stimulate color changes in the epidermis. The receptors have been

isolated are under study. Some speculate that it is involved in sleep, biorhythms, or pigmentation

(note indicator of primary adrenal insufficiency below). MSH should not be confused with

melatonin, a hormone produced by the pineal gland that stimulates lightening of skin in fish and

amphibians and whose function in humans is also not well understood. Altered secretion of MSH

has been implicated in causing skin pigmentation during the menstrual cycle and pregnancy. The

two major types of melanotropin, α-MSH and β-MSH, are derived from ACTH and β-lipotropin,

respectively.

α-MSH contains the same amino acid sequence as the first 13 amino acids of ACTH; β-MSH has

Protein and Polypeptide Hormones 17

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

18 amino acid residues. A third melanotropin, γ- melanotropin, is derived from a larger peptide

precursor, pro-opiomelanocortin (POMC).

Pro-opiomelanocortin (POMC) production is stimulated by Corticotropin-releasing hormone

(CRH) of the hypothalamus. POMC is an ultra-cool 260 amino acid protein, which gets

glycosylated (sugars added, probably at select arginine residues as for prolactin) and then

cleaved to give a number of neurohormones. The anterior pituitary hormones, adenocorticotropin

hormone (ACTH) and LPH (lipotropin hormone) are derived from POMC in the pars distalis of

the anterior pituitary. In the pars intermedia area of the anterior pituitary, the ACTH derived

from cleavage of POMC is cleaved further to produce α-MSH (melanocyte-stimulating

hormone). Thus, POMC is processed differently in different areas of the pituitary. We can also

get β-endorphin out of POMC cleavage, which means POMC is a source of an endogenous opiod

peptide. In fact, POMC is the precursor to ACTH,

α,β,γ-MSH, α,β,γ-endorphin, β-lipoprotein, and met-enkephalin. Besides the hypophysis

(pituitary), the brain, gastrointestinal system, immune cells, placenta, and gonads are able to

synthesize POMC products.

Lipotropins (Enkephalins and Endorphins) The isolation of two peptides with opiate like

activity from pig brains was a breakthrough in the year 1975. These related pentapeptides, called

methionine-enkephalin (Fig. 5) and leucine-enkephalin (Fig. 6), are abundant in certain

terminals and have been shown to occur in the pituitary gland.

1 5 1 5
Tyr-Gly-Gly-Phe-Met Tyr-Gly-Gly-Phe-Leu
[Met]Enkephalin [Leu]Enkephalin

Fig. 5 Fig. 6

Protein and Polypeptide Hormones 18

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

The structure of enkephalins revealed that the amino acid sequence of met-enkephalin is

identical with the sequence of residues 61-65 of β-lipotropin (β-LPF), a larger peptide found in

the pituitary gland. This discovery suggested that β-LPH might be a precursor for other larger

peptides containing the met-enkephalin sequence. Soon after the structural relationship between

met-enkephalin and β-LPH was established, longer peptides, called endorphins, were isolated

from the intermediate lobe of the pituitary gland.

The endorphins (α, β and γ) contained the met-enkephalinamino acid sequence and possessed

morphine like activity. -endorphin, the longest of these peptides, is a 31 amino acid

endogenous opiod peptide most important to the neuroendocrine regulation of human

reproduction. Co-released with ACTH from the pituitary, -endorphin is also produced in the

medial-basal hypothalamus and widely distributed in the brain. Of the three major types of opiod

receptors, , , and  receptors, -endorphin selectively binds  receptors, the same that binds

morphine with high affinity. Opiod  receptors are mostly localized to the limbic system and

hypothalamus.

Opiod receptors regulate or modulate functions other than pain including temperature perception,

hunger and thirst control, and reproduction. Only in the presence of steroids (such as estrogen or

testosterone) can -endorphin inhibit GnRH release from the mediobasal hypothalamus and

consequently LH (leutinizing hormone) by the anterior pituitary. LH is important for ovulation.

The endorphins and enkephalins have a wide range of biological activities and most of their

activities are in the CNS. They inhibit the release of dopamine in the brain tissue and

acetylcholine from the neuromuscular junctions.

Protein and Polypeptide Hormones 19

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

Growth Hormone (Somatotropin)

Human placental lactogen (hPL), GH, and prolactin (PRL) comprise the growth hormone family.

All have about 200 amino acids, 2 disulfide bonds, and no glycosylation. Although each has

special receptors and unique characteristics to their activity, they all possess growth-promoting

and lactogenic activity. Mature GH (22,000 daltons) is synthesized in acidophilic pituitary

somatotropes as a single polypeptide chain. Because of alternate RNA splicing, a small amount

of a somewhat smaller molecular form is also secreted.

While details of the method of signal transduction by the members of the GH family of tropic

hormones remain unclear, PKC activity has been demonstrated to correlate directly with the

biological effects of PRL and GH. This appears to indicate that the PKC signal transduction

pathway is operative in transducing signals for the GH family of hormones.

The role of growth hormone in regulating IGF-1 production was noted above. Humans respond

to natural or recombinant human or primate growth hormone with appropriate secretion of IGF-

1, but growth hormone of other species has no normal biological effect in man. The latter is

puzzling because interspecies GH homologies are quite high in many cases, and most other

species respond well to human growth hormone. In humans, growth hormone promotes

gluconeogenesis and is consequently hyperglycemic. It promotes amino acid uptake by cells,

with the result that GH therapy puts an organism into positive nitrogen balance, similar to that

seen in growing children. Finally, growth hormone is lipolytic, inducing the breakdown of tissue

lipids and thus providing energy supplies that are used to support the stimulated protein synthesis

induced by increased amino acid uptake.

There are a number of genetic deficiencies associated with GH. GH-deficient dwarfs lack the

ability to synthesize or secrete GH, and these short-statured individuals respond well to GH

Protein and Polypeptide Hormones 20

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

therapy. Pygmies lack the IGF-1 response to GH but not its metabolic effects; thus in pygmies

the deficiency is post-receptor in nature. Finally, Laron dwarfs have normal or excess plasma

GH, but lack liver GH receptors and have low levels of circulating IGF-1. The defect in these

individuals is clearly related to an inability to respond to GH by the production of IGF-1. The

production of excessive amounts of GH before epiphyseal closure of the long bones leads to

gigantism, and when GH becomes excessive after epiphyseal closure, acral bone growth leads to

the characteristic features of acromegaly.

Prolactin (PRL)

Prolactin is produced by acidophilic pituitary lactotropes. Prolactin is the lone tropic hormone of

the pituitary that is routinely under negative control by prolactin inhibiting hormone (PIH),

which is now known to be dopamine. Decreased hypophyseal dopamine production, or damage

to the hypophyseal stalk, leads to rapid up-regulation of PRL secretion. A number of other

hypothalamic releasing hormones induce increased prolactin secretion; as a result, it is unclear

whether a specific PRH exists for up-regulating PRL secretion. PRL initiates and maintains

lactation in mammals, but normally only in mammary tissue that has been primed with

estrogenic sex hormones.

Placental Hormones

Human Chorionic Gonadotropin

Human chorionic gonadotropin (hCG) is a glycoprotein synthesized by the placenta. Estrogens

stimulate the anterior pituitary to produce placentotropin, which in turn stimulates hCG synthesis

and secretion. It exerts effects that are similar to those of pituitary LH.

Protein and Polypeptide Hormones 21

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

hCG is used therapeutically in the management of cryptorchidism in prepubertal boys. It is also

used in women in conjunction with menotropins to induce ovulation when the endogenous

availability of gonadotropin is not normal.

Human Placental Lactogen (hPL)

Human placental lactogen is produced by the placenta late in gestation. At its height it is secreted

at a rate of about 1 g/day, the highest secretory rate of any known human hormone. However,

little hPL reaches the fetal circulation, and hPL has only about 1% the activity of PRL or GH in

producing biological effects, leading some to question its functional importance in humans. It

has been identified as a protein composed of 191 amino acid units in a single-peptide chain with

two disulfide bridges. hPL resembles human somatotropin.

Gonadotropic Hormones: Follicle Stimulating Hormone (FSH), Luteinizing Hormone

(LH), Chorionic Gonadotropin (CG)

FSH, LH, and CG is each glycosylated to the extent of 16-30% of their molecular weight. each

hormone is composed of two chains with combined molecular weights ranging from MW 30-

38,000.The 89 amino acid alpha-chain for FSH, LH, and CG (89 + 3 N-terminal AAs) is

esentially identical for each. The beta-chains, then, differ to confer selectivity with regards to

their actions. For FSH and LH, the beta- chains have 115 AA, for CG the beta-chain has 145 AA.

As in the case of TSH, the chains are held together by non-covalent forces (ionic, hydrogen

bond, Van der Waals but not disulfide bonds). Within each chain, there are dissulfide bonds, 5 in

the alpha- and 6 in the beta-chains.

Protein and Polypeptide Hormones 22

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

Functions: FSH, LH, and CG have different effects on women and men (not surprising), but the

effects are consistent with the preparation for and development of the fertilized ova.

Follicle Stimulating Hormone (FSH)

Females:

A. FSH promotes the development and maturation of ovarian follicles (many develop

simultaneously, but at different rates)

B. FSH stimulates estrogen synthesis and secretion by ovarian follicles. In follicles grown in

tissue culture, addition of exogenous FSH causes estrogen secretion to increase at a rate

much faster than development stimulation, indicating that the effect on estrogen level is

independent of and not merely the consequence of increased follicle development.

Males:

A. FSH is essential for gametogenesis and sperm formation.

Luteinizing Hormone (LH)

In women, one function of LH is that it induces ovulation in the ovary (FSH alone cannot induce

ovulation). At first, LH contributes to the final maturation and development of both the follicle

and the ovum which it contains. After a period of time, a sharp increase in blood plasma estradiol

(estrogen), a positive feedback to follicle development, is followed a day later by a near

simultaneous release of FSH and LH from the anterior pituitary. In response to the burst of FSH

and LH, one or more mature follicles rupture and release an ovum (egg).

Protein and Polypeptide Hormones 23

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

A second function of LH in women is that it acts upon the ruptured follicle to stimulate its

conversion to a corpus luteum, which provides an ovarian source of estrogen and subsequently

progesterone. Progesterone exerts a negative feedback to depress the production of LH and hence

prevent further follicle and ovum development until progesterone levels subside at the end of the

menstrual cycle. If no postive stimulus to the corpus luteum is received from placental CG

(chorionic gonadotropin) within seven days, the corpus luteum begins to recede or degenerate.

As the corpus luteum degenerates, it stops secreting estrogen and progesterone, which then leads

to a reinitiation of the cycle. The cycle begins again because low levels of estrogen and

progesterone stimulate the anterior pituitary to increase its output of FSH and LH.

In men, LH stimulates the formation and secretion of androgens, especially testosterone, by the

testes.

Chorionic Gonadotropin (CG)

CG is secreted by the chorion (the outermost extraembryonic membrane which gives rise to the

placenta) seven (7) days after ovulation if the ovum has been fertilized (i.e. in pregnancy). CG

stimulates the ovarian corpus luteum to secrete high levels of estrogen and progesterone.

The α (alpha) chains of FSH, LH, and CH are almost identical, as mentioned above, and are quite

similar to the α (alpha) chain of TSH (thyroid stimulatory hormone). The differences in activity

are caused by differences in the β (beta) chains. There is an 82% homology in AA 1-115 between

the b chains of LH and CG. Individually, that is in the monomeric rather than dimeric state, the

separated chains for any of the gonadotropins show no activity. However, due to the near

identical nature of the α chains, separation of the α chain from one gonadotropin and reassembly

Protein and Polypeptide Hormones 24

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

with the b chain of another gonadotropin produces a fully active hormone with the activity as

specified by the β chain.

Each chain contains carbohydrate. The alpha-chains each have an Asn linked oligosaccharide. In

the FSH beta-chain, there are two carbohydrates each Asn linked. The LH beta-chain contains

one Asn linked carbohydrate. CG contains a total of six (6) carbohydrate chains, two (2) Asn-

linked in a region similar to that of FSH and four (4) linked through Ser in a non-homologous

region to FSH or LH.

Carbohydrate sidechains, as seen for PRL and TSH, are often found on proteins which are

excreted and may contribute to stability or recognition. Such biological effects are the

consequence of the carbohydrate's structure. The carbohydrates contribute complexities to the

spacial structure which do not occur with nonglycosylated proteins. Carbohydrates are made up

of numerous linked sugars. They can branch (at mannose) and offer multiple possibilities for

formation of hydrogen bonds, esters, and ethers through use of the hydroxyl (-OH) groups which

stick up or down around the sugar ring. Through such bonding interactions, particularly the non-

covalent hydrogen bonding, they can be involved in the recognition of receptors. As an example

of carbohydrate protection of the peptide backbone from enzymatic degradation, removal of one

carbohydrate from FSH changes its half life from 90 minutes to 2-3 minutes.

A single DNA gene encodes for the protein which is the alpha-chain of FSH, LH, CG, and TSH.

While the alpha-chain proteins are nearly identical, differences between them must be the

Protein and Polypeptide Hormones 25

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

consequence of alternative messenger RNA splicing or post-translational enzyme processing,

such as alternative proteolysis or glycosylation.

The genes for the gonadotropin beta-chains are found on different chromosomes. While there are

seven (7) genes that code for human CG (chorionic gonadotropin), only two appear to be

transcribed (made from DNA into RNA) and translated (RNA to protein). The physiological

impact of different forms for CG has yet to be evaluated.

The affinities for FSH, LH, and CG for their receptors is very high with a KD of ~10-10 M. The

receptors exist in excess, so a receptor reserve exists. One gets maximal response when only 1%

of the hormone receptors are occupied.

Prolonged exposure of receptors to hormones leads to down regulation of receptors. This

decrease in the number of gonadotropin receptors in the plasma membrane of ovary or corpus

luteum cells can result from either an internalization (endocytosis) of existing receptors, a

decrease in their rate of synthesis or both. Due to the receptor reserve and the low percentage of

receptor occupation needed for response, considerable down regulation of receptors is necessary

before noticeable effects are observable.

The removal of the carbohydrate from a gonadotropin does not affect the binding affinity per se,

but does markedly reduce the abilities of the hormones to stimulate adenylate cyclase or

steroidogenesis. Deglycosylated hormones can act as a competitive antagonist. So FSH without

carbohydrate can compete with FSH with carbohydrate.

Thyroid Stimulating Hormone (TSH)

Secretion of TSH (also called thyrotropin), the final member of the glycoprotein hormone

family, is stimulated by thyrotropin-releasing hormone, TRH from the hypothalamus. While

Protein and Polypeptide Hormones 26

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

cAMP causes increased secretion of TSH by thyrotropes, it is not yet certain that cAMP is the

physiological signal that regulates TSH production.

Circulating TSH binds to receptors on the basal membrane of thyroid follicles. The receptors are

coupled through a G-protein to adenylate cyclase. The result is that ligand binding increases

thyrocyte cAMP and PKA, leading in the short term to increased secretion of thyroxin (T4) and

triiodothyronine (T3). Chronic stimulation of the receptor causes an increase in the synthesis of a

major thyroid hormone precursor, thyroglobulin.

Thyroglobulin produced on rough endoplasmic reticulum has a molecular weight of 660,000. It

is glycosylated and contains more than 100 tyrosine residues, which become iodinated and are

used to synthesize T3 and T4. Thyroglobulin is exoctosed through the apical membrane into the

closed lumen of thyroid follicles, where it accumulates as the major protein of the thyroid and

where maturation takes place. Briefly, a Na+/K+-ATPase-driven pump concentrates iodide (I-) in

thyroid cells, and the iodide is transported to the follicle lumen. There it is oxidized to I + by a

thyroperoxidase found only in thyroid tissue. The addition of I + to tyrosine residues of

thyroglobulin is catalyzed by the same enzyme, leading to the production of thyroglobulin

containing monoiodotyrosyl (MIT) and diiodotyrosyl (DIT) residues. The thyronines, T3 and T4,

are formed by combining MIT and DIT residues on thyroglobulin.

Mature, iodinated thyroglobulin is taken up in vesicles by thyrocytes and fuses with lysosomes.

Lysosomal proteases degrade thyroglobulin releasing amino acids and T3 and T4, which are

secreted into the circulation. These compounds are very hydrophobic and require a carrier

protein for delivery to target tissues. In the plasma, T3 and T4 are bound to a carrier glycoprotein

known as thyroxin-binding globulin and are disseminated throughout the body in this form.

Protein and Polypeptide Hormones 27

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

Thyroid hormones act by binding to cytosolic receptors very similar to steroid hormone

receptors, and for this reason T3 and T4 are often classified along with the hydrophobic steroid

hormones. The principal role of thyroid hormones is also like that of steroid hormones. In adults,

the ligand receptor combination binds to thyroid hormone response elements in nuclear DNA

and is responsible for up-regulating general protein synthesis and inducing a state of positive

nitrogen balance. In the embryo, thyroid hormone is necessary for normal development.

Hypothyroidism in the embryo is responsible for cretinism, which is characterized by multiple

congenital defects and mental retardation.

Thyroid stimulating autoantibodies (TSAb) also activate the human thyroid TSH receptor,

leading to the hyperthyroidism of Graves' disease. TSAbs bind to the TSH receptor and mimic

the TSH stimulation of the gland by increasing intracellular cAMP.

The feedback loop that regulates T3 and T4 production is a single short negative loop, with the

T3 and T4 being responsible for down-regulating pituitary TSH secretion. Meanwhile,

continuously secreted hypothalamic TRH is responsible for up-regulating TSH production. The

TSH actually secreted by thyrotropes is the net result of the negative effects of T3 and T4 and the

positive effect of TRH.

Somatostatin

Somatostatin is an oligopeptide (14 amino acid residues) and is referred to as somatotropin

release-inhibiting factor (SRIF) (Fig.7). It acts by both endocrine and paracrine pathways to

affect its target cells. A majority of the circulating somatostatin appears to come from the

pancreas and gastrointestinal tract.

Protein and Polypeptide Hormones 28

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

S S
1 5 10
Ala-Gly-Cys-Lys-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-Cys

Fig. 7

If one had to summarize the effects of somatostatin in one phrase, it would be: "somatostatin

inhibits the secretion of many other hormones".

Effects on the Pituitary Gland

Somatostatin was named for its effect of inhibiting secretion of growth hormone from the

pituitary gland. Experimentally, all known stimuli for growth hormone secretion are suppressed

by somatostatin administration. Additionally, animals treated with antiserum to somatostatin

show elevated blood concentrations of growth hormone, as do animals that are genetically

engineered to disrupt their somatostatin gene.

Ultimately, growth hormone secretion is controlled by the interaction of somatostatin and growth

hormone releasing hormone, both of which are secreted by hypothalamic neurons.

Effects on the Pancreas

Cells within pancreatic islets secrete insulin, glucagon and somatostatin. Somatostatin appears to

act primarily in a paracrine manner to inhibit the secretion of both insulin and glucagon. It also

has the effect in suppressing pancreatic exocrine secretions, by inhibiting cholecystokinin-

stimulated enzyme secretion and secretin-stimulated bicarbonate secretion.

Effects on the Gastrointestinal Tract

Somatostatin is secreted by scattered cells in the GI epithelium, and by neurons in the enteric

nervous system. It has been shown to inhibit secretion of many of the other GI hormones,

including gastrin, cholecystokinin, secretin and vasoactive intestinal peptide.

Protein and Polypeptide Hormones 29

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

In addition to the direct effects of inhibiting secretion of other GI hormones, somatostatin has a

variety of other inhibitory effects on the GI tract, which may reflect its effects on other

hormones, plus some additional direct effects. Somatostatin suppresses secretion of gastric acid

and pepsin, lowers the rate of gastric emptying, and reduces smooth muscle contractions and

blood flow within the intestine. Collectively, these activities seem to have the overall effect of

decreasing the rate of nutrient absorption.

Effects on the Nervous System

Somatostatin is often referred to as having neuromodulatory activity within the central nervous

sytem, and appears to have a variety of complex effects on neural transmission. Injection of

somatostatin into the brain of rodents leads to such things as increased arousal and decreased

sleep, and impairment of some motor responses.

Pharmacological Uses

Somatostatin and its synthetic analogs are used clinically to treat a variety of neoplasms. It is

also used in to treat giantism and acromegaly, due to its ability to inhibit growth hormone

secretion.

Pancreatic Hormones: Insulin and Glucagon

The pancreas mainly produces two hormones, insulin and glucagon. Insulin is secreted by the β-

cells and glucagon by the α-cells. The primary function of the pancreatic hormones is the

regulation of whole-body energy metabolism, principally by regulating the concentration and

activity of numerous enzymes involved in catabolism and anabolism of the major cell energy

supplies.

Protein and Polypeptide Hormones 30

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

Insulin The earliest of the pancreatic hormones recognized was insulin, whose major function is

to counter the concerted action of a number of hyperglycemia-generating hormones and to

maintain low blood glucose levels. Because there are numerous hyperglycemic hormones,

untreated disorders associated with insulin generally lead to severe hyperglycemia and shortened

life span. Insulin is a member of a family of structurally and functionally similar molecules that

include the insulin-like growth factors (IGF-1 and IGF-2), and relaxin. The tertiary structure of

all 4 molecules is similar, and all have growth-promoting activities, but the dominant role of

insulin is metabolic while the dominant roles of the IGFs and relaxin are in the regulation of cell

growth and differentiation.

Proinsulin, which is a single-chain of 86 amino acid residues, is enzymatically transformed into

insulin within the β-cells. The conversion involves the cleavage of a connecting C-peptide,

which contains between 30-35 residues, the number and sequence varying among different

species; human C-peptide consists of 35 residues. The resulting human insulin consists of two

peptide chains, designated A (having 21 residues) and B (having 30 residues), which are inter-

chain connected by two disulfide bonds. Furthermore, the A-chain also contains an intra-chain

disulfide bond between Cys6 and Cys11.

Actions of Insulin

The major function of insulin is to counter the concerted action of a number of hyperglycemia-

generating hormones and to maintain low blood glucose levels. Because there are numerous

hyperglycemic hormones, untreated disorders associated with insulin generally lead to severe

hyperglycemia and shortened life span.

Protein and Polypeptide Hormones 31

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

In addition to its role in regulating glucose metabolism, insulin stimulates lipogenesis,

diminishes lipolysis, and increases amino acid transport into cells. Insulin also modulates

transcription, altering the cell content of numerous mRNAs. It stimulates growth, DNA

synthesis, and cell replication, effects that it holds in common with the insulin-like growth

factors (IGFs) and relaxin.

Insulin, secreted by the β-cells of the pancreas, is directly infused via the portal vein to the liver,

where it exerts profound metabolic effects. These effects are the response of the activation of the

insulin receptor, which belongs to the class of cell surface receptors that exhibit intrinsic tyrosine

kinase activity. The insulin receptor is a heterotetramer of 2 extracellular α-subunits disulfide

bonded to 2 transmembrane β-subunits. With respect to hepatic glucose homeostasis, the effects

of insulin receptor activation are specific phosphorylation events that lead to an increase in the

storage of glucose with a concomitant decrease in hepatic glucose release to the circulation.

In most nonhepatic tissues, insulin increases glucose uptake by increasing the number of plasma

membrane glucose transporters: GLUTs. Glucose transporters are in a continuous state of

turnover. Increases in the plasma membrane content of transporters stem from an increase in the

rate of recruitment of new transporters into the plasma membrane, deriving from a special pool

of preformed transporters localized in the cytoplasm. GLUT1 is present in most tissues, GLUT2

is found in liver and pancreatic b-cells, GLUT3 is in the brain and GLUT4 is found in heart,

adipose tissue and skeletal muscle.

Protein and Polypeptide Hormones 32

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

In liver glucose uptake is dramatically increased because of increased activity of the enzymes

glucokinase, phosphofructokinase-1 (PFK-1), and pyruvate kinase (PK), the key regulatory

enzymes of glycolysis. The latter effects are induced by insulin-dependent activation of

phosphodiesterase, with decreased PKA activity and diminished phosphorylation of pyruvate

kinase and phosphofructokinase-2, PFK-2. Dephosphorylation of pyruvate kinase increases its'

activity while dephosphorylation of PFK-2 renders it active as a kinase. The kinase activity of

PFK-2 converts fructose-6-phosphate into fructose-2,6-bisphosphate (F2,6BP). F2, 6BP is a

potent allosteric activator of the rate limiting enzyme of glycolysis, PFK-1, and an inhibitor of

the gluconeogenic enzyme, fructose-1,6-bisphosphatase. In addition, phosphatases specific for

the phosphorylated forms of the glycolytic enzymes increase in activity under the influence of

insulin. All these events lead to conversion of the glycolytic enzymes to their active forms and

consequently a significant increase in glycolysis. In addition, glucose-6-phosphatase activity is

down regulated. The net effect is an increase in the content of hepatocyte glucose and its

phosphorylated derivatives, with diminished blood glucose.

In addition to the above described events, diminished cAMP and elevated phosphatase activity

combine to convert glycogen phosphorylase to its inactive form and glycogen synthase to its

active form, with the result that not only is glucose funneled to glycolytic products, but glycogen

content is increased as well.

Insulin also has profound effects on the transcription of numerous genes, effects that are

primarily mediated by regulated function of sterol-regulated element binding protein, SREBP.

These transcriptional effects include (but are not limited to) increases in glucokinase, pyruvate

Protein and Polypeptide Hormones 33

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

kinase, lipoprotein lipase (LPL), fatty acid synthase (FAS) and acetylCoA carboxylase (ACC)

and decreases in glucose 6-phosphatase, fructose 1,6-bisphosphatase and phosphoenolpyruvate

carboxykinase (PEPCK).

Glucagon is a 29-amino acid straight chain polypeptide (Fig. 8) synthesized by the α-cells of the

Islets of Langerhans as a very much larger proglucagon molecule. Like insulin, glucagon lacks a

plasma carrier protein, and like insulin its circulating half-life is also about 5 minutes. As a

consequence of the latter trait, the principal effect of glucagon is on the liver, which is the first

tissue perfused by blood containing pancreatic secretions. The role of glucagon is well

established. It binds to plasma membrane receptors and is coupled through G-proteins to

adenylate cyclase. The resultant increases in cAMP and PKA reverse all of the effects described

above that insulin has on liver. The increases also lead to a marked elevation of circulating

glucose, with the glucose being derived from liver gluconeogenesis and liver glycogenolysis.

1 5 10

His-Ser-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr
20 15
Gln-Ala-Arg-Arg-Ser-Asp-Leu-Tyr-Lys-Ser
25

Asp-Phe-Val-Gln-Tyr-Leu-Met-Asn-Thr

Fig. 8

Gastrointestinal Hormones and Peptides

There are more than 30 peptides currently identified as being expressed within the digestive

tract, making the gut, the largest endocrine organ in the body. The regulatory peptides

synthesized by the gut include hormones, peptide neurotransmitters and growth factors. As a

Protein and Polypeptide Hormones 34

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

matter of fact, several hormones and neurotransmitters first identified in the central nervous

system and other endocrine organs have subsequently been found in endocrine cells and/or

neurons of the gut.

The gastrointestinal hormones and peptides have significant physiological roles. These are

tabulated below (Table 2).

Table 2

Hormone Location Major Action

Enteroendocrine L cells Potentiates glucose-dependent
Glucagon-like peptide 1
predominantly in the insulin secretion, inhibits glucagon
(GLP-1)
ileum and colon secretion, inhibits gastric emptying
Glucose-dependent
insulinotropic polypeptide Enteroendocrine K cells
Inhibits secretion of gastric acid,
(GIP) originally called of the duodenum and
enhances insulin secretion
gastric inhibitory proximal jejunum
polypeptide
Gastrin (17-residue
Gastric antrum,
polupeptide) Gastric acid and pepsin secretion
duodenum
(Fig. 9)
Cholecystokinin-
Stimulates gallbladder contraction
Pancreozymin
Duodenum, jejunum and bile flow, increases secretion of
(CCK-PZ) (33-residue
digestive enzymes from pancreas
polypeptide) (Fig. 10)
Secretin (27-amino acid
Duodenum, jejunum Pancreatic bicarbonate secretion
polypeptide) (Fig. 11)
Vasoactive intestinal Smooth muscle relaxation;
peptide (VIP) (28-residue Pancreas stimulates pancreatic bicarbonate
polypeptide) (Fig. 12) secretion

Protein and Polypeptide Hormones 35

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

Motilin (22-residue Initiates interdigestive intestinal

Small bowel
polypeptide) motility
Pancreatic polypeptide Inhibits pancreatic bicarbonate and
Pancreas
(PP) protein secretion
Stomach, duodenum,
Enkephalins Opiate-like actions
gallbladder
CNS function in pain (nociception),
involved in vomit reflex, stimulates
Entire gastrointestinal
Substance P salivary secretions, induces
tract
vasodilatation antagonists have anti-
depressant properties
Bombesin-like Stimulates release of gastrin and
Stomach, duodenum
immunoreactivity (BLI) CCK
Causes vasodilatation, increases
Neurotensin (13-amino vascular permeation and gastrin
Ileal mucosa
acid peptide) secretion, decreases gastric acid and
secretin secretion

1 5 10
Lys-Ala-Pro-Ser-Gly-Arg-Val-Ser-Met-Ile
1 5 20 15
(pyro)Glu-Gly-Pro-Trp-Met His-Ser-Pro-Asp-Leu-Ser-Gln-Leu-Asn-Lys
10 25 30
Ala-Glu-Glu-Glu-Glu-Glu Arg-Ile-Ser-Asp-Arg-Asp-Tyr-Met-Gly-Trp
15

HO3S Tyr-Gly-Trp-Met-Asp-Phe NH2 SO3H

Fig. 9 H2N Phe-Asp-Met

Cholecystokinin

Fig. 10

Protein and Polypeptide Hormones 36

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

1 5 10 1 5 10
His-Ser-Asp-Gly-Thr-Phe-Thr-Ser-Gly-Leu His-Ser-Asp-Ala-Val-Phe-Thr-Asp-Asn-Tyr
20 15 20 15
Gln-Leu-Arg-Ala-Ser-Asp-Arg-Leu-Arg-Ser Lys-Val-Ala-Met-Gln-Lys-Arg-Leu-Arg-Thr
25
25
Arg-Leu-Leu-Gln-Gly-Leu-Val-NH2 Lys-Tyr-Leu-Asn-Ser-Ile-Leu-Asn

Fig. 11 Fig. 12

Neurophyseal Hormones: Vasopressin and Oxytocin

The principal hormones of the posterior pituitary are the nonapeptides oxytocin and vasopressin.

These substances are synthesized as prohormones in neural cell bodies of the hypothalamus and

mature as they pass down axons in association with carrier proteins termed neurophysins. The

axons terminate in the posterior pituitary, and the hormones are secreted directly into the

systemic circulation.

Vasopressin is also known as antidiuretic hormone (ADH) (Fig. 13), because it is the main

regulator of body fluid osmolarity. The secretion of vasopressin is regulated in the hypothalamus

by osmoreceptors, which sense water concentration and stimulate increased vasopressin

secretion when plasma osmolarity increases. The secreted vasopressin increases the reabsorption

rate of water in kidney tubule cells, causing the excretion of urine that is concentrated in Na + and

thus yielding a net drop in osmolarity of body fluids. Vasopressin deficiency leads to watery

urine and polydipsia, a condition known as diabetes insipidus. Vasopressin binds plasma

membrane receptors and acts through G-proteins to activate the cAMP/PKA regulatory system.

Protein and Polypeptide Hormones 37

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

S S
1 5
NH2-Cys-Tyr-Phe-Gln-Asn-Cys-Pro-Arg-Gly-NH2

Fig. 13

Oxytocin

The mechanism of action of oxytocin (Fig. 14) is unknown. Oxytocin secretion in nursing

women is stimulated by direct neural feedback obtained by stimulation of the nipple during

suckling. Its physiological effects include the contraction of mammary gland myoepithelial cells,

which induces the ejection of milk from mammary glands, and the stimulation of uterine smooth

muscle contraction leading to childbirth.

S S
1 5
NH2-Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Leu-Gly-NH2

Fig. 14

Parathyroid Hormone (PTH)

Parathyroid hormone (PTH, molecular weight 9,500) is synthesized and secreted by chief cells of

the parathyroid in response to systemic Ca2+ levels. The Ca2+ receptor of the parathyroid gland

responds to Ca2+ by increasing intracellular levels of PKC, Ca2+ and IP3; this stage is followed,

after a period of protein synthesis, by PTH secretion. The synthesis and secretion of PTH in chief

cells is constitutive, but Ca2+ regulates the level of PTH in chief cells (and thus its secretion) by

increasing the rate of PTH proteolysis when plasma Ca2+ levels rise and by decreasing the

proteolysis of PTH when Ca2+ levels fall. The role of PTH is to regulate Ca2+ concentration in

extracellular fluids. The feedback loop that regulates PTH secretion therefore involves the

parathyroids, Ca2+, and the target tissues described below.

Protein and Polypeptide Hormones 38

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

PTH acts by binding to cAMP-coupled plasma membrane receptors, initiating a cascade of

reactions that culminates in the biological response. The body response to PTH is complex but is

aimed in all tissues at increasing Ca2+ levels in extracellular fluids. PTH induces the dissolution

of bone by stimulating osteoclast activity, which leads to elevated plasma Ca 2+ and phosphate. In

the kidney, PTH reduces renal Ca2+ clearance by stimulating its reabsorption; at the same time,

PTH reduces the reabsorption of phosphate and thereby increases its clearance. Finally, PTH acts

on the liver, kidney, and intestine to stimulate the production of the steroid hormone 1,25-

dihydroxycholecalciferol (calcitriol), which is responsible for Ca 2+ absorption in the intestine.

Calcitonin (CT, Thyrocalcitonin)

Calcitonin (CT) (Fig. 15) is a 32-amino acid peptide secreted by the parafollicular cells of the

thyroid gland in response to hypocalcemia. Calcitonins as obtained from different species are

identical at 7 of the first 9 residues, contain Gly at position 28, and all terminate with Pro-NH2.

The C-terminal proline amide (Pro-NH2) is very important for the biologic function of CT, as is

the disulfide-bridge between Cys residues at positions 1 and 7. Calcitonin is employed

therapeutically to relieve the symptoms of osteoporosis, although details of its mechanism of

action remain unclear. However, it has been observed that CT induces the synthesis of PTH in

isolated cells, which leads in vivo to increased plasma Ca2+ levels. In addition, CT has been

shown to reduce the synthesis of osteoporin (Opn), a protein made by osteoclasts and responsible

for attaching osteoclasts to bone. Thus, is appears that CT elevates plasma Ca 2+ via PTH

induction and reduces bone reabsorption by decreasing osteoclast binding to bone.

Protein and Polypeptide Hormones 39

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

1
Cys S S Cys Met
25
Gly Thr Leu Gln Thr
5 10
Asn Ser Gly Pro Ala
Leu
Thr Phe Ile

Tyr Thr Gly

20
Thr His Val
30
Gln Phe Gly
15
Asp Lys Ala

Phe Asn Pro-NH2

Fig. 15

Renin-Angiotensin System

The renin-angiotensin system is responsible for regulation of blood pressure. The intrarenal

baroreceptor system is a key mechanism for regulating renin secretion. A drop in pressure results

in the release of renin from the juxtaglomerular cells of the kidneys. Renin secretion is also

regulated by the rate of Na+ and Cl- transport across the macula densa. Higher the rate of

transport of these ions, lower is the rate of renin secretion. The only function for renin is to

cleave a 10-amino acid peptide from the N-terminal end of angiotensinogen (Fig. 16). This

decapeptide is called angiotensin I. Angiotensin I is then cleaved by the action of angiotensin-

converting enzyme, ACE to the active hormone, angiotensin II (Fig. 16), which is an

octapeptide. ACE removes 2 amino acids from the C-terminal end of angiotensin I. Angiotensin

II is hydrolyzed to angiotensin III, a heptapeptide, by an aminopeptidase. Angiotensin III

preserves most of the pharmacological activities of its precursor. Further degradation of

angiotensin III leads to pharmacologically inactive peptide fragments.

Protein and Polypeptide Hormones 40

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

1 1
Asp Phe His Asp Phe His
10 10
Arg Pro Leu α - Globulin Arg Pro Leu
2 Renin

Val His Val Ser Val His

5 5
Tyr Ile Ile Tyr Ile
His
Angiotensinogen Angiotensin I

ACE

1
Asp Phe

Arg Pro
Val His
5
Tyr Ile
Angiotensin II

Fig. 16

Angiotensin II was also referred to as hypertensin and angiotonin. It is one of the most potent

naturally occurring vasoconstrictors. The vasoconstrictive action of angiotensin II is primarily

exerted on the arterioles and leads to a rise in both systolic and diastolic blood pressure. In

individuals that are depleted of sodium or who have liver disease (e.g. cirrhosis), the pressive

actions of angiotensin II are greatly reduced. These conditions lead to increased circulating levels

of angiotensin II which in turn leads to a down-regulation in the number of angiotensin II

receptors on smooth muscle cells. As a consequence, administration of exogenous angiotensin II

to these individuals has little effect.

Other physiological responses to angiotensin II include induction of adrenal cortex synthesis and

secretion of aldosterone. Angiotensin II also acts on the brain leading to increased blood

pressure, vasopressin and ACTH secretion and increased water intake. Angiotensin II affects the

Protein and Polypeptide Hormones 41

Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

contractility of the mesangial cells of the kidney leading to decreased glomerular filtration rate.

One additional effect of angiotensin II is to potentiate the release of norepinephrine.

Two distinct types of angiotensin II receptors have been identified, AT 1 and AT2. The AT1

receptors are classical serpentine (7 transmembrane spanning) receptors. The AT 1 receptors are

coupled to a G-protein that leads to activation of PLC-γ. Although the AT2 receptors are also

serpentine, they do not appear to be coupled to activation of G-proteins.

Natriuretic Hormones

Natriuresis refers to enhanced urinary excretion of sodium. This can occur in certain disease

states and through the action of diuretic drugs. At least 3 natriuretic hormones have been

identified. Atrial natiuretic peptide (ANP) was the first cardiac natriuretic hormone identified.

This hormone is secreted by cardiac muscle when sodium chloride intake is increased and when

the volume of the extracellular fluid expands. Active ANP is a 28-amino acid peptide containing

a 17-amino acid ring formed by intrachain disulfide bonding. Two smaller forms of ANP have

also been isolated from the brain. A brain natriuretic peptide (BNP) (first isolated from porcine

brain) has been identified and found in human heart and blood (but not human brain). BNP has

different amino acids in its 17-amino acid ring and is encoding for by a different gene. In

humans, a third natriuretic peptide (CNP) is present in the brain but not in the heart.

The action of ANP is to cause natriuesis presumably by increasing glomerular filtration rate (its

exact mechanism of action remains unclear). ANP induces relaxation of the mesangial cells of

the glomeruli and thus may increase the surface area of these cells so that filtration is increased.

Alternatively, ANP might act on tubule cells to increase sodium excretion. Other effects of ANP

Protein and Polypeptide Hormones 42

 Pharmacy e-Book Medicinal Chemistry Dr. Asif Husain

include reducing blood pressure, decreasing the responsiveness of adrenal glomerulosa cells to

stimuli that result in aldostreone production and secretion, inhibit secretion of vasopressin and

decreasing vascular smooth muscle cell responses to vasoconstrictive agents. These latter actions

of ANP are counter to the effects of angiotensin II. In fact, ANP also lowers renin secretion by

the kidneys thus lowering circulating angiotensin II levels.

Three different ANP receptors have been identified: ANPR-A, ANPR-B and ANPR-C. When

ANP, BNP or CNP bind to receptor, an increase in guanylate cyclase activity results leading to

production of cyclic GMP (cGMP). Both ANPR-A and ANPR-B proteins span the plasma

membrane and their intracellular domains possess intrinsic guanylate cyclase activity. The exact

function of the ANPR-C protein is unclear as this receptor does not contian an intracellular

domain with intrinsic guanylylate cyclase activity. It is hypothesized that it may act through a G-

protein that activates PLC- and inhibits adenylate cyclase or that it acts simply as a clearance

receptor removing natriuretic peptides from the blood.

Protein and Polypeptide Hormones 43

View publication stats