Medicinal case 2

Statins
Fatima 1210552
Hiba 1212867
Sara 1212210
Israa 1201441
Introduction
Statins are an important type of medicine used to lower cholesterol levels by blocking
certain enzymes. They are among the most widely used cholesterol-lowering drugs and
make up a large part of the pharmaceutical market. These drugs not only help improve
health but also bring big profits to the companies that manufacture them. In 2002,
atorvastatin and simvastatin earned about $7 billion and $5.3 billion, respectively.
In this study, we will look at how statins were discovered, how they work in the body, and
how they help reduce cholesterol and prevent heart disease.
Cholesterol and coronary heart disease
Cholesterol is crucial for cell membrane function and steroid hormone
biosynthesis. Excessive cholesterol can lead to cardiovascular disease.
Cholesterol is transported through low-density lipoprotein (LDL) or high-density
lipoprotein (HDL) particles. LDLs are 22 nm in diameter and have a mass of 3
million Daltons, containing a lipoprotein of 4536 amino acid residues, linoleate,
phospholipids, and cholesterol molecules. They transport cholesterol and
triglycerides from the liver to peripheral tissues. HDLs, 8-11 nm in diameter, carry
fatty acids and cholesterol back to the liver. High levels of LDL or low levels of HDL
can lead to mortality from coronary heart disease. Retained cholesterol can lead to
fatty plaques, narrowing arteries, and increased risk of atherosclerosis. Lowering
LDL levels and increasing HDL can reduce the risk of heart attacks and strokes.
The target enzyme
 The target enzyme
In order the drug to work effectively, it must target the most important enzyme that catalyses
the rate-limiting step in the interaction. The target enzyme here is HMG-CoA reductase, this
enzyme is responsible for the rate-limiting step in cholesterol biosynthesis. Converting HMG-
CoA into mevalonate using NADPH as a cofactor.
 HMG-CoA reductase and binding
HMG-CoA reductase is composed of four protein subunits with two active sites located between these
subunits. It uses specific amino acids to stabilize the substrate (HMG-CoA) and facilitate the
reaction:

1. Lys-735 forms an ionic bond with the carboxylate group of HMG-CoA.

2. Ser-684 and Asp-690 form hydrogen bonds with alcohol groups.
3. Lys-691 forms hydrogen and ionic bonds, stabilizing both the intermediate (mevaldyl-CoA)
and the transition state.
 The catalytic reaction; 1st step:

The catalytic reaction occurs in two steps, each involving a hydride transfer from NADPH:

1. HMG-CoA is converted to mevaldyl-CoA, with His-866 donating a proton to release

coenzyme A.
The catalytic reaction; 2nd step:

2. Mevaldyl-CoA is reduced to mevalonate, with Glu-559 donating proton. The enzyme’s

structure ensures stabilization of intermediates and reduces the activation energy.
HMG-CoA reductase inhibition
HMG-CoA reductase plays a crucial role in regulating cholesterol production in the body. It is
highly regulated to maintain cholesterol balance. Its activity decreases in response to high
cholesterol levels through:
● Phosphorylation (inactivation),
● Reduced transcription and translation, and
● Increased degradation.
Statins
Statins, a class of cholesterol-lowering drugs, work by mimicking the enzyme's natural
substrate, competitively inhibiting its activity, but do not undergo the enzymatic reaction.
Statins interact strongly with the enzyme due to:
● Polar and hydrophobic interactions.
● Flexibility of the enzyme, which creates a binding site for the statins’ hydrophobic
regions.

This reduces cholesterol synthesis and effectively lowers LDL (bad) cholesterol levels in
the blood.
The discovery of statins
 The Discovery of Compactin (Mevastatin)
The First Statin: Compactin

Discovered in the 1970s by Akira Endo from Penicillium citrinum.

Compactin was the first inhibitor of HMG-CoA reductase.

Mechanism: Mimics HMG-CoA to competitively inhibit the enzyme.

Structure:

Lactone ring: Hydrolyzed to an active form.

Decalin ring: Hydrophobic interactions with the enzyme.

The Development of mevinolin (Lovastatin)
Lovastatin: The First Commercial Statin

Isolated by Merck & Co. from Aspergillus terreus in 1978.

Approved for clinical use in 1987 as the first cholesterol-lowering statin.

Improved on compactin’s structure, enhancing safety and efficacy.

Pioneered a new class of cholesterol-lowering drugs.

Evolution and Impact of Statins
The Statin Revolution : Statins evolved into two main types:

● Type II Statins: Fully synthetic, with improved

● Type I Statins: Natural or semi- potency and safety (e.g., Atorvastatin,
synthetic (e.g., Lovastatin, Simvastatin) Rosuvastatin).

● Clinical Impact:
Atorvastatin (Lipitor) became the world’s best-selling
drug.
Statins have significantly reduced cardiovascular risks
globally
Mechanism of action for statins-
pharmacodynamics
 Mechanism of action for statins-pharmacodynamics

The statins work by acting as competitive inhibitors . They mimic the natural substrate and
compete with it in order to bind to the active site.

Unlike the natural substrate, they do not undergo an enzyme- catalysed reaction and they
bind more strongly
How can we explain all of this?

Both the polar head group and the hydrophobic moieties are important to the action of statins. All
the statins share the same polar head-group and it is this group which mimics the natural
substrate (HMG-SCoA).

The head-group of the statins can, therefore, mimic the natural substrate and bind to the active
site using the same binding interactions. We now need to explain why statins bind more strongly
than the natural substrate and why they are resistant to the enzyme-catalysed reaction.

the statins contain an extra hydrophobic region which can form additional hydrophobic interactions
with a hydrophobic binding region present in the enzyme. This allows the statins to bind more
strongly.
the statins are resistant to the enzyme- catalysed reaction as the coenzyme A moiety in the about statins

They are actually more similar to the first intermediate in the enzyme-catalysed mechanism—mevaldyl
CoA— than to the substrate

Assuming that mevaldyl CoA is less stable than the substrate, this implies that the statins bear
some resemblance to the transition state leading to mevaldyl CoA. Consequently, they would be
expected to have a stronger binding interaction than the natural substrate and are likely to be
acting as transition state analogues .
Binding interactions of statins
X-ray crystallography studies the binding interactions of substrates and statins.
The polar head-group of statins binds similarly to the substrate, while the
hydrophobic region of coenzyme A is narrow and cannot accommodate the bulky
hydrophobic groups present in statins. The statins' binding to the enzyme reflects
the enzyme's flexibility, as it creates a lower hydrophobic binding region next to the
active site that can accommodate the hydrophobic moiety present in the statin.
This makes statins effective inhibitors, as they can take advantage of the
enzyme's flexibility and create their own binding site.
The binding interactions of type I and type II statins with the enzyme reveal that
type II statins bind to the same hydrophobic region as type I statins, with additional
interactions with amino acids like leucine, valine, and alanine. A crucial interaction
involves the fluorophenyl group and an arginine residue, allowing additional
interactions. Atorvastatin and rosuvastatin form extra hydrogen bonding
interactions with the enzyme, enhancing their efficacy in clearing LDL cholesterol
from the plasma.
Looking at Figure CS1.12, the arginine residue in the binding region firstly has a polar interaction between
this residue and the fluorine substituent and secondly the planar guanidinium group of the residue is stacked
on the phenyl ring allowing for additional interactions. Atorvastatin and rosuvastatin can form additional
hydrogen bonding interactions with the enzyme that do not occur with other statins. This involves a serine
residue that acts as a hydrogen bond donor to the carbonyl oxygen atom of atorvastatin (Figure CS1.12)

FIGURE
CS1.12 Binding interactions for the hydrophobic

moiety of atorvastatin with HMGR (3-hydroxy-3-

methylglutaryl-coenzyme A reductase).
Other mechanisms of action for statins
Beyond Cholesterol Reduction
Statins have pleiotropic effects that enhance cardiovascular protection.

 Upregulation of LDL Receptors:

● Increases liver uptake of LDL cholesterol, lowering blood levels.

 Anti-Inflammatory Effects:
● Reduces inflammatory markers like C-reactive protein (CRP).
● Improves endothelial function by boosting nitric oxide production.
Reducing Atherosclerotic Risk
Protecting Against Plaques and Damage

Improved Endothelial Function:

 Antioxidant Properties:
● Lowers oxidative stress and oxidized LDL, reducing plaque formation.
● Decreases production of reactive oxygen species (ROS).

 Plaque Stabilization:
● Reduces plaque lipid content and strengthens fibrous caps.
● Stabilized plaques lower the risk of heart attacks and strokes.

 Emerging Benefits:
● Supports bone health and exhibits anti-thrombotic effects.
Other targets for cholesterol lowering drugs
 Other targets for cholesterol lowering drugs
enzymes are involved in cholesterol biosynthesis. The early attempts to find cholesterol-lowering drugs
focused on inhibiting enzymes in the later steps of this biosynthesis process.

Why Later Steps Matter: Inhibiting enzymes in the later steps of biosynthesis is more selective because it
reduces cholesterol levels without affecting the production of other compounds that share part of the same
biosynthetic pathway.

inhibitors in the later steps were effective, they caused an accumulation of unused substrates, which were
insoluble and toxic.

Statins as a Solution: When HMG-CoA reductase is inhibited by statins, the accumulated substrate is water-
soluble and can be easily metabolized, preventing toxic buildup.

Combination with Other Drugs: Statins are also used alongside drugs that target proteins not directly involved
in cholesterol biosynthesis. For example, Vytorin combines simvastatin and ezetimibe, which reduces
cholesterol absorption in the gastrointestinal tract.
Question
• The conversion of Carbutamide to Tolbutamide is an example of one of the
lead modification methods.

1) Explain why Carbutamide has both hypoglycemic and antibacterial

activity?
2) What is the advantage of the modified compound Tolbutamid
Answer:

Q1: Carbutamide is a sulfonamide derivative with the following structural features:

Hypoglycemic activity: The sulfonylurea group (–SO2–NH–CO–NH–) interacts with ATP-sensitive potassium (K_ATP)
channels in pancreatic beta cells. By closing these channels, the compound promotes insulin release, thereby reducing
blood glucose levels.
Antibacterial activity: The presence of the sulfonamide group (–SO2–NH2), a structural analog of para-aminobenzoic
acid (PABA), interferes with bacterial folate synthesis. This disrupts nucleotide synthesis and bacterial replication,
resulting in antibacterial effects. Thus, Carbutamide’s structure enables it to act both as a hypoglycemic agent (via
interaction with K_ATP channels) and as an antibacterial agent (by inhibiting bacterial folate synthesis).

Q2: The conversion of Carbutamide to Tolbutamide involves replacing the primary amine group (–NH2) with a methyl
group (–CH3) in the para position of the aromatic ring. This structural modification has the following effects:
Improved selectivity for hypoglycemic activity: The removal of the primary amine group eliminates antibacterial
activity, reducing undesired side effects related to interference with bacterial processes.
Better pharmacokinetic properties: The addition of the methyl group improves the compound’s lipophilicity, leading to
better absorption, distribution, and bioavailability.
Focused therapeutic action: Tolbutamide is more selective for K_ATP channels in pancreatic beta cells, enhancing its
effectiveness as a hypoglycemic agent while minimizing off-target actions.Overall, Tolbutamide represents a more
refined drug with reduced side effects and enhanced efficacy for managing diabetes. This modification illustrates the lead
optimization process in drug development, where structural changes are made to enhance desired activity and minimize
undesired properties.