Patrick: An Introduction

to Medicinal Chemistry 5e
Case Study 1

STATINS AS
ANTI-CHOLESTEROL AGENTS
1. Cholesterol

H H
HO

Notes
Important in biosynthesis and cell membrane structure
Excess cholesterol leads to cardiovascular disease
Fatty molecule transported round blood supply by low-density
and high-density lipoproteins (LDLs and HDLs)
LDLs carry cholesterol to cells
HDLs carry cholesterol from cells to liver
Mortality is associated with high levels of LDLs or low levels of
HDLs
Cholesterol can cause fatty plaques in arteries leading to a risk
of artherosclerosis, clot formation, stroke and heart attack
2. Target for statins
Target enzyme

O HO CH3 O HO CH3 O
HMG-CoA reductase
+ 2H + CS
oH
A
CoA S O HO O

(S)-HMG-CoA 2NADPH 2NADP (R)-Meval onate

Notes on statins
Cholesterol is synthesised within the cell
One way of lowering cholesterol levels in the blood is to block its
synthesis (more than 30 enzymes involved)
Inhibit an enzyme in the biosynthetic pathway to cholesterol
Prevent synthesis of cholesterol within cells
Target the enzyme catalysing the rate limiting step in the
biosynthetic pathway (enzyme catalyse reduction step)
Cholesterol can still be obtained from the diet (diet control)
3. Catalytic mechanism

O H O O H OH
NADPH +H NADPH + H
R S CoA R S CoA R H NADP R H
NADP
Mevaldyl-CoA Mevaldehyde
+ CoA
SH

Notes
Involves two hydride transfers
Two molecules of cofactor required (NADPH)
3. Catalytic mechanism

Substrate binding

Asp-690
Lys-691 Ser-684
N O O
H H O
H H-bond H
H H-bond
H-bond O O CH3 O
ionic
CoA S O
H3N Lys-735
3. Catalytic mechanism

His-866
Lys-691 Lys-691 Lys-691
N N His-866 N N NH
H H H H H H
H H Ionic H
H-bondO H-bond H N NH
H-bond O
O
SH CoA
R S CoA R S CoA R H
H
Mevaldyl-CoA Mev aldehyde
H
(fromNADPH)

Notes
Lys, His, Glu and Asp are involved in the reaction mechanism
Histidine acts as an acid catalyst
Lysine stabilises the negatively charged oxygen of mevaldyl-CoA and the
transition state leading to it
Lowers activation energy for first step
3. Catalytic mechanism
Glu-559
Lys-691 Lys-691 Glu-559
Lys-691
N O O N
N H H H H O O
H H H Ionic H H
H
H-bond
O H-bond OH
H-bondO
R H
R H H R H
H
Mevalonate
H
(fromNADPH)

Notes
Glutamic acid acts as an acid catalyst (unusual unionized ) due to aspartate
role.
An aspartate residue stabilises Glu-559 and Lys-691
O Asp-767
Lys-691 Glu-559
N O
H H H
H O O
Mechanism of the first reduction

Mechanism of the second reduction

4. Identification of a Lead Compound

HO O

O
O H Compactin (Mevastatin)
O IC50 = 23 nM
H

Notes
Identified by screening compounds produced by microorganisms
Microbes lacking HMGR might produce HMGR inhibitors to inhibit
microbes which do have HMGR - chemical warfare (competition)
Compactin (Mevastatin) isolated from Penicillium citrinum
10,000 x higher affinity for enzyme than substrate
Never reached the market (toxic effects were observed during preclinical
trials)
 5. Type I Statins
HO O HO O HO
CO2H
O O OH
O H O H O H
O O O
H H Me H H

HO
Lovastatin Simvastatin Pravastatin
IC50 = 24 nM IC50 = 24 nM IC50 = 1900 nM

Notes
Lovastatin isolated from Aspergillus terreus
First statin to be marketed (Merck; 1987) compactin
Revolutionised treatment of hypercholesterolaemia
Simvastatin introduced in 1988 as semi-synthetic analogue of lovastatin
Pravastatin derived from compactin by biological transformation (1991)
5. Type I Statins
HO
CO2H
Polar head
OH
O H
O
R H
R'
Hydrophobic
moiety
R''
Decalin ring

Notes
General structure of type I statins contains a polar head and a hydrophobic
moiety including a decalin ring
Lovastatin and simvastatin are prodrugs where a lactone ring is hydrolysed
to give the polar head
5. Type I Statins
HO
CO2H
* Polar head
* OH
O H
* O
R H
R'
= asymmetric centres * * ** Hydrophobic
* moiety
*
R''
Decalin ring

Disadvantages of Type I statins

Various side effects
Difficult to synthesise
Large number of asymmetric centres
6. Type II Statins
HO HO HO
CO2H CO2H CO2H
OH OH OH
H H H
F
F F
N
N
O N
NH H3C
O

Fluvastatin Atorvastatin Cerivastatin

IC50 = 28 nM IC50 = 8 nM IC50 = 10 nM
Notes
Synthetic agents
Contain larger hydrophobic moiety with no asymmetric centres
Easier to synthesise
Fluvostatin (1994), atorvastatin (1997), cerivastatin (1998),
rosuvastatin (2003)
6. Type II Statins
HO HO
CO2H CO2H
OH OH
H H
F F

N N N
Rosuvastatin Pitavastatin
IC50 = 5 nM H3C N IC50 = 6.8 nM
S CH3
O
O

Notes
Structures share a number of similar features (me better drugs)
Rosuvastatin is the most potent - related to sulfonamide group
Cerivastatin is the most hydrophobic
Pravastatin and rosuvastatin are the least hydrophobic
6. Type II Statins

Notes
Statins with lower hydrophobic character target liver cells and
have lower side effects
Less hydrophobic statins do not cross cell membranes easily
Liver cells have transport proteins for statins whereas other cells
do not
Majority of cholesterol synthesis takes place in liver cells
Side effects thought to be due to inhibition of HMGR in other
cells such as muscle cells
Common side effect is myalgia (muscle pain)
Rhabdomyolysis = severe muscle toxicity which can be fatal
Cerivastatin withdrawn in 2001 due to rhabdomyolysis and 50
fatalities
7. Statins - Mechanism of action
H CH3
HO O HO CH3 O HO
CO2 CO2
OH CoA S O = O
H HMG-SCoA
Hydrophobic group SCoA
Statins

Notes
Competitive inhibitors of HMGR
Polar head group mimics the natural substrate (HMG-SCoA)
Same binding interactions for polar head group as natural
substrate
Hydrophobic moiety forms additional binding interactions
Binds more strongly than natural substrate, but does not
undergo reaction - no leaving group
7. Statins - Mechanism of action

CH3 H
HO HO
CO2 CO2
OH OH
H H
SCoA Hydrophobic group

Me v a ldy l C oA St a t ins

Notes
Statins are closer mimics of the first reaction intermediate
mevaldyl CoA
Statins likely to bear a resemblance to the transition state for the
first stage of the reaction mechanism
Can be viewed as transition-state analogues
8. Statins - Binding interactions
OH
HO2C
HO
H Ion-dipole
bond Arg-590
Leu HN
v dw F
Val N
Ala HN
NH2

HN
O
Atorvastatin H-bond
H
O

Notes Ser-565
Polar head group binds in similar manner to substrate
Hydrophobic moiety does not bind to the pocket for SCoA
Enzyme is flexible and alters shape to accommodate statins
Hydrophobic pocket is created to bind the hydrophobic moiety
8. Statins - Binding interactions
OH
HO2C
HO
methylethyl H
substituent Ion-dipole
bond Arg-590
Leu v dw HN
F
Val N
Ala HN
NH2

HN
O
Atorvastatin H-bond
H
O

Ser-565

The methylethyl substituent of Type II statins binds to the same

region of the binding site as the decalin ring of type I statins
8. Statins - Binding interactions
OH
HO2C
HO fluorophenyl
H substituent Ion-dipole
bond Arg-590
Leu HN
v dw F
Val N
Ala HN
NH2

HN
O
Atorvastatin H-bond
H
O

Ser-565

Arg-590 forms an important polar interaction with the

fluorophenyl substituent
The planar guanidium group is also stacked over the phenyl
ring
8. Statins - Binding interactions
OH
HO2C
HO
H Ion-dipole
bond Arg-590
Leu HN
v dw F
Val N
Ala HN
NH2

amide group HN
O
Atorvastatin H-bond
H
O

Ser-565
Amide group forms an additional hydrogen bonding interaction
with Ser-565
Additional interaction not formed with other statins other than
rosuvastatin
8. Statins - Binding interactions
OH
HO2C
HO
Ion-dipole
H bond Arg-590
Leu F HN
v dw
Val
Ala HN
NH2

N N
Rosuvastatin N CH3
H3C S
O H
O H-bond O
sulfone group
Ser-565

Notes
Rosuvastatin forms additional H-bonding interactions
Sulfone oxygen forms a hydrogen bonding interaction with Ser 565
Sulfone group also interacts uniquely with Arg-568
Explains why rosuvastatin is the most potent statin
Sulfone group important for binding as well as for selectivity
9. Other mechanisms of action for statins

Notes
Statins inhibit HMGR in liver cells
HMGR inhibition lowers cholesterol levels in liver cells
Causes an increase in the synthesis of hepatic LDL receptors
Increases the number of LDL receptors in the cell membrane of
liver cells (cell requires cholesterol it produces LDL receptors)
Increases the amount of LDL-cholesterol cleared from the
plasma
Crucial to the effectiveness of statins