Lipoprotein metabolism

PSC 3110 Fall 2004

Lipoproteins
particles found in plasma that transport
lipids including cholesterol
lipoprotein classes
chylomicrons: take lipids from small intestine
through lymph cells
very low density lipoproteins (VLDL)
intermediate density lipoproteins (IDL)
low density lipoproteins (LDL)
high density lipoproteins (HDL)
Lipoprotein
class
Density
(g/mL)
Diameter
(nm)
Protein %
of dry wt
Phosphol
ipid %
Triacylglycerol
% of dry wt
HDL 1.063-1.21 5 15 33 29 8
LDL 1.019
1.063
18 28 25 21 4
IDL 1.006-1.019 25 - 50 18 22 31
VLDL 0.95 1.006 30 - 80 10 18 50
chylomicrons < 0.95 100 - 500 1 - 2 7 84
Composition and properties of human lipoproteins
most proteins have densities of about 1.3 1.4 g/mL and lipid aggregates usually
have densities of about 0.8 g/mL
Lipoprotein structure
LDL molecule
The apolipoproteins
major components of lipoproteins
often referred to as aproteins
classified by alphabetical designation (A thru
E)
the use of roman numeral suffix describes the
order in which the apolipoprotein emerge from
a chromatographic column
responsible for recognition of particle by
receptors
HELICAL WHEEL PROJECTION OF A
PORTION OF APOLIPOPROTEIN A-I
LIPOPROTEINS
spherical particles with a hydrophobic
core (TG and esterified cholesterol)
apolipoproteins on the surface
large: apoB (b-48 and B-100) atherogenic
smaller: apoA-I, apoC-II, apoE
classified on the basis of density and
electrophoretic mobility (VLDL; LDL;
IDL;HDL; Lp(a)
Apoproteins of human
lipoproteins
A-I (28,300)- principal protein in HDL
90 120 mg% in plasma; activates LCAT
A-II (8,700) occurs as dimer mainly in HDL
30 50 mg %; enhances hepatic lipase activity
B-48 (240,000) found only in chylomicron
<5 mg %; derived from apo-B-100 gene by RNA
editing; lacks the LDL receptor-binding domain of
apo-B-100
B-100 (500,000) principal protein in LDL
80 100 mg %; binds to LDL receptor
C-I (7,000) found in chylomicron, VLDL, HDL
4 7 mg %; may also activate LCAT
C-II (8,800) - found in chylomicron, VLDL, HDL
3 8 mg %; activates lipoprotein lipase
C-III (8,800) - found in chylomicron, VLDL, IDL, HDL
8 15 mg %; inhibits lipoprotein lipase
D (32,500) - found in HDL
8 10 mg %; also called cholesterol ester transfer protein (CETP)
E (34,100) - found in chylomicron, VLDL, IDL HDL
3 6 mg %; binds to LDL receptor
H (50,000) found in chylomicron; also known as b-2-
glycoprotein I (involved in TG metabolism)


Apoproteins of human
lipoproteins
Major lipoprotein classes
Chylomicrons (derived from diet)
density <<1.006
diameter 80 - 500 nm
dietary triglycerides
apoB-48, apoA-I, apoA-II, apoA-IV, apoC-
II/C-III, apoE
remains at origin in electrophoretic field
Chylomicron
formed through extrusion of resynthesized
triglycerides from the mucosal cells into the
intestinal lacteals
flow through the thoracic ducts into the
suclavian veins
degraded to remnants by the action of
lipoprotein lipase (LpL) which is located on
capillary endothelial cell surface
remnants are taken up by liver parenchymal
cells due to apoE-III and apoE-IV isoform
recognition sites
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Major lipoprotein classes
VLDL
density >1.006
diameter 30 - 80nm
endogenous triglycerides
apoB-100, apoE, apoC-II/C-III
prebeta in electrophoresis
formed in the liver as nascent VLDL
(contains only triglycerides, apoE and apoB)

VLDL
nascent VLDLs then interact with HDL
to generate mature VLDLs (with added
cholesterol, apoC-II and apoC-III)
mature VLDLs are acted upon by LpL to
generate VLDL remnants (IDL)
IDL are further degraded by hepatic
triglyceride lipase (HTGL) to generate
LDLs
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Major lipoprotein classes
IDL (intermediate density lipoproteins)
density: 1.006 - 1.019
diameter: 25 - 35nm
cholesteryl esters and triglycerides
apoB-100, apoE, apoC-II/C-III
slow pre-beta
Major lipoprotein classes
LDL (low density lipoproteins)
density: 1.019 - 1.063
diameter: 18-25nm
cholesteryl esters
apoB-100
beta (electrophoresis)
< 130 LDL cholesterol is desirable, 130-159
is borderline high and >160 is high
Major lipoprotein classes
HDL (high density lipoproteins)
density: 1.063-1.210
diameter: 5-12nm
cholesteryl esters and phospholipids
apoA-I, apoA-II, apoC-II/C-III and apoE
alpha (electrophoresis)
HDLs
Several subfamilies exist
Discoidal HDL :
contains cholesterol, phospholipid, apoA-I, apoA-
II, apoE and is disc shaped;
it is formed in liver and intestine
It interacts with chylomicra remnants and
lecithin-cholesterol acyl transferase (LCAT) to
form HDL
3

HDLs
HDL3
composed of cholesterol, cholesterol ester, phospholipid
and apoA and apoE
interacts with the cell plasma membranes to remove free
cholesterol
reaction with LCAT converts HDL
3
to HDL
2a
(an HDL
with a high apoE and cholesterol ester content)
cholesterol ester-rich HDL
2a
is then converted to
triglyceride-rich HDL
2b
by concomitant transfer of HDL
cholesterol esters to VLDL and VLDL triglycerides to HDL




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Functions of HDL
transfers proteins to other lipoproteins
picks up lipids from other lipoproteins
picks up cholesterol from cell membranes
converts cholesterol to cholesterol esters via the
LCAT reaction
transfers cholesterol esters to other
lipoproteins, which transport them to the liver
(referred to as reverse cholesterol transport)
Lipoproteins (a)- Lp(a)
another atherogenic family of lipoproteins(at
least 19 different alleles)
they consist of LDL and a protein designated as
(a)
the apoA is covalently linked to apoB-100 by a
disulfide linkage
unusual in that it contains a kringle protein
motif/domain (tri-looped structure with 3
intramolecular disulfide bonds resembling a
Danish pretzel)
high risk association with premature coronary
artery disease and stroke
Cholesterol and lipid transport by
lipoproteins
Cholesterol and lipid transport by
lipoproteins
The LDL receptor
characterized by Michael Brown and Joseph
Goldstein (Nobel prize winners in 1985)
based on work on familial
hypercholesterolemia
receptor also called B/E receptor because of its
ability to recognize particles containing both
apos B and E
activity occurs mainly in the liver
receptor recognizes apo E more readily than
apo B-100
Representation of the
LDL receptor (839 aa)

extracellular domain is
responsible for apo-B-
100/apo-E binding

intracellular domain is
responsible for
clustering of LDL
receptors into coated
pit region of plasma
membrane
Cholesterol sources, biosynthesis
and degradation
diet
only found in animal fat
biosynthesis
primarily synthesized in the liver from acetyl CoA
biosynthesis is inhibited by LDL uptake by the liver
degradation
only occurs in the liver
cholesterol is converted to bile acids
Biosynthesis of cholesterol
- synthesis of acetoacetyl CoA
Biosynthesis of cholesterol
- synthesis of mevalonate
rate-limiting step
and step subject to
inhibition by statins
Biosynthesis of cholesterol
-synthesis of isopentenyl
-pyrophosphate
A monoterpene
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Biosynthesis of cholesterol
- synthesis of squalene
a sesquiterpene
a triterpene
S
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o
f

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Biosynthesis of cholesterol
- synthesis of lanosterol
the allylamine antifungals
interfere with the epoxidation
step (naftidine, terfinabine)
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Biosynthesis of cholesterol
The demethylation
of lanosterol is also
a useful step for drug
design i.e. azole antifungals
ACAT inhibitors act
here
Biosynthesis summary
Bile acids from cholesterol
formed from cholesterol in the liver
stored in the gall bladder in bile as bile
salts (sodium and potassium)
utilized during digestion of fats and other
lipid substances (act as detergents)
rate limiting step is the conversion of
cholesterol to 7-alpha cholesterol by 7-a-
hydroxylase
HO
HO OH
7a-hydroxylase
NADPH + H
+
NADP
cholesterol
7a-hydroxycholesterol
HO OH
OH
C S
O
CoA
H
HO OH
C S
O
CoA
H
chenodeoxycholyl- CoA
cholyl-CoA
12a-hydroxylase
O
2
; NADPH + H
+
2 CoA-SH
O
2
NADPH + H
+
2 CoA-SH
HO

C S
O
CoA
H
cholyl-CoA
HO

C
O
H
N CH
2
COOH
glycine
CoA-SH
Conversion of cholyl-CoA to
glycocholic acid
glycocholic acid
HO

C S
O
CoA
H
cholyl-CoA
HO

C
O
H

CH
2
CH
2
SO
3
H
H
taurocholic acid (primary bile acid)
taurine
CoA-SH
Conversion of
cholyl CoA to
taurocholic acid
Taurine
Taurine is formed by the decarboxylation
of cysteic acid, which in turn is made by
oxidation of cysteine
C
COO
H H
3
N
CH
2
SH
C
COO
H H
3
N
CH
2
SO
2
cysteine
cysteine sulfinate
CH
2
H
3
N
CH
2
SO
3
taurine
+ O
2 + O
2
- CO
2
HO OH
C S
O
CoA
H
chenodeoxycholyl- CoA
tauro- and glyco-chenodeoxycholic acids
(primary bile acids)
HO
H
COOH
lithocholic acid (secondary bile acid)
deconjugation +
7a-dehydrxylation
(catalyzed by microbial
enzymes)
HO

C
O
H
N CH
2
COOH
glycocholic acid (primary bile acid)
HO

H
COOH
deconjugation +
7
a
-dehydroxylation

catalyzed by microbial
enzymes)
Deoxycholic acid (secondary bile acid
Conversion of glycocholic
acid to deoxycholic acid
Bile acids
cholic acid is the bile acid found in the
largest amount in bile
cholic acid and chenodeoxycholic acid are
referred to as primary bile acids
bile acids are converted to either glycine
or taurine conjugates (in humans the
ratio of glycine to taurine conjugates is
3:1)

Approximate composition of bile salts
Glycocholate 24%
Glycochenodeoxycholate 24%
Taurocholate 12%
Taurochenodeoxycholate 12%
Glycodeoxycholate- 16%
Taurodeoxycholate 8%
Various lithocholate 4%

Bile acids
fat digestion products are absorbed in the first
100 cm of small intestine
the primary and secondary bile acids are
reabsorbed almost exclusively in the ileum
returning to the liver by way of the portal
circulation (98 to 99%)
this is known as the enterohepatic circulation
less than 500 mg a day escapes reabsorption
and is excreted in the feces
Bile salts
detergent character of bile salts is due to
the hydrophobic-hydrophilic nature of
the molecules
the presence of hydroxyl (or sulfate) and
the terminal carboxyl group on the tail
gives the molecule its hydrophilic face
the steroid ring with its puckered plane
provides the hydrophobic face
Function of bile salts
emulsification of fats due to detergent activity
aid in the absorption of fat-soluble vitamins
(especially vitamin K)
accelerate the action of pancreatic lipase
have choleretic action stimulate the liver to
secrete bile
stimulate intestinal motility
keep cholesterol in solution (as micelles)
Mixed micelle formed by bile salts, triacylglycerols andf pancreatic
lipase
CH
3
CH
3
CH
3
COOH
H
H
H
H
CHOLANIC ACID
CH
3
CH
3
CH
3
COOH
H
H
H
H
HO
OH
HO
CH
3
CH
3
CH
3
COOH
H
H
H
H
HO
OH
CH
3
CH
3
CH
3
COOH
H
H
H
H
HO
OH
CHOLIC ACID
CHENODEOXYCHOLIC ACID (CHENODIOL)
URSODEOXYCHOLIC ACID (URSODIOL)
(ACTIGALL)
(CHENIX)
BILE ACIDS
3 7
12
GALLSTONE THERAPEUTIC AGENTS
chenodeoxycholic acid (chenodiol; Chenix)
ursodeoxycholic acid (ursodiol; Actigall)
MAO:
reduce hepatic secretion of cholesterol into
bile
inhibition of HMGCoA reductase: inhibit
cholesterol biosynthesis
increase cholesterol solubility
Chenodiol and ursodiol
both are effective in dissolving cholesterol
stones in some patients
ursodiol is the 7-beta epimer of chenodiol
most effective in dissolving small (<5
mm) floating stones in a functioning
gallbladder
cannot dissolve stones that are more than
4% calcium by weight
Atherosclerosis
hardening of the arteries due to the
deposition of atheromas
heart disease is the leading cause of death
caused by the deposition of cholesteryl
esters on the walls of arteries
atherosclerosis is correlated with high
LDL and low HDL
Photograph of an arterial plaque
Frederickson -WHO classification
Type I: incr. chylomicrons, reduced HDL, absence of
lipoprotein lipase; deficiency of apo CII
(hyperchylomironemia)
Type II-A: raised LDL; decreased catabolism of LDL
(receptor deficiency or polygenic)
Type II-B: raised VLDL + LDL; often reduced HDL;
increased production of VLDL + impaired LDL
catabolism
Type III: raised IDL (dysbetalipoproteinemia); abnormal
apolipoprotein E; impaired catabolism of IDL; elevated
cholesterol and triglycerides (formerly known as broad
beta disease)

Frederickson -WHO classification
Type IV: raised VLDL; often reduced HDL;
impaired VLDL catabolism; dietary
indiscretion ( formerly known as
hyperprebetalipoproteinemia)

Type V: raised chylomicrons + VLDL; reduced
HDL; reduced lipoprotein lipase + VLDL
hypersecretion (formerly known as mixed
lipemia)
Factors promoting elevated blood
lipids
age
men >45 years of age; women > 55 years of age
family history of CAD
smoking
hypertension >140/90 mm Hg
low HDL cholesterol
obesity >30% overweight
diabetes mellitus
inactivity/ lack of exercise
Mechanisms of action of drugs
bind to bile acids/cholesterol
inhibit absorption/reabsorption
increase peroxisomal FA oxidation
stimulate lipoprotein lipase
inhibit triglyceride lipase
inhibit HMG CoA reductase
stimulates microsomal 7-alpha
hydroxylase
Drug Classification
systemic/non-sytemic
cholesterol lowering agents
bile acid sequestrants
sitosterols*
probucol*
d-thyroxin*
HMG Co-A reductase inhibitors
* No longer available commercially in the U.S
Drug Classification
mixed activity (nicotinic acid)
triglyceride lowering
clofibrate (Atromid-S)
gemfibrosil (Lopid)
fenofibrate (Tricor)
Bile sequestering resins
H
C
H
2
C
H
C
H
2
C
CH
H
2
C
N(CH
3
)
3
n
CHOLESTYRAMINE
H
2
N
HN
HN
HN
(CH
2
)
6
N(CH
3
)
3
HN
(CH
2
)
9
-CH
3
OH
HN
(CH
2
)
6
N(CH
3
)
3
HN
(CH
2
)
9
CH
3
H
2
N
. n HCl
. n HCl
. n HCl
. n HCl
. n HCl
. n HCl
. n HCl
. n HCl
COLESEVELAM
Bile sequestering resins
CH
2
HN
CH
2
CH
2
N CH
2
CH
2
N
CH
2
CH
2
CH
2
COH COH COH H
CH
2
CH
2
N CH
2
CH
2
COH
CH
2
H H
CH
2
N
CH
2
H
N N CH
2
CH
2
-CH
2
CH
2
N
CH
2
NH
CH
2
COH
CH
2
N H CH
2
CH
2
H CH
2
-CH
2
-
n
COLESTIPOL (COLESTID)
Bile acid sequestrants
po, safest, non systemic
bind to bile acids and inhibit reabsorption
increase 7-alpha hydroxylase activity leading to
cholesterol degradation
decreases plasma LDL
problems:
abdominal discomfort, bloating, constipation
decreases drug absorption; wait 4 hrs after
administration of BAS to give drugs
Colesevelam (WelChol)
polyalkylamine hydrochloride) cross linked
with epichlorohydrin and alkylated with 1-
bromodecane and (6-bromohexyl)
trimethylammonium bromide
available as a 625 mg tablet
same mechanism of action as colestipol and
cholestyramine
Bile sequestering resins
drug interactions
(decreased serum
level)
aspirin
clindamycin
clofibrate
furosemide
glipzide
tolbutamide
phenytoin
imipramine
methyldopa
nicotinic acid
penicillin G
propranolol
tetracycline
thiazide diuretics
digoxin
hydrocortisone
phosphate supplements
CH
3
CH
3
H
3
C
CH
2
CH
3
CH
3
CH
3
HO
STIGMASTANOL
CH
3
CH
3
H
3
C
CH
2
CH
3
CH
3
CH
3
HO
BETA SITOSTEROL
PLANT STEROLS
CH
3
CH
3
H
3
C
CH
2
CH
3
CH
3
CH
3
HO
STIGMASTEROL
CH
3
CH
3
H
3
C
CH
3
CH
3
HO
CH
3
CAMPESTROL
CH
3
CH
3
H
3
C
CH
3
CH
3
HO
CH
3
CH
3
ALPHA1-SITOSTEROL
More plant sterols
HMG CoA reductase
3 different regulatory mechanisms are
involved:
covalent modification: phosphorylation by
cAMP-dependent protein kinases inactivate the
reductase. This inactivation can be reversed by 2
specific phosphatases
degradation of the enzyme half life of 3 hours
and the half-life depends on cholesterol levels
gene expression: cholesterol levels control the
amount of mRNA
O
COOH
CH
3
OH
SCoA
OH
COOH
CH
3
OH
H H
2 NADPH
2 NADP
- CoASH
OH
COOH
CH
3
OH
H
SCoA
O
COOH
CH
3
OH
H
NADPH
NADPH
O
CH
3
H
3
C
O
H
3
C
O
H
H
H
O
CH
3
H
3
C
CH
3
O
H
HO
CH3
N
P
O
COOH
Ph
O-
HO
Et
F
O
O
H
HO
Cl
Cl HO
Ki = 10
-10
Ki = 10
-11
Ki = 10
-10
Lovastatin
CH
3
H
O
O
HO O
H H
3
C
O
MEVASTATIN
CH
3
H
O
O
HO O
H H
3
C
H
3
C
O
LOVASTATIN (MEVACOR)
CH
3
H
O
O
HO O
CH
3
H
3
C
O
SIMVASTATIN (ZOCOR)
CH
3
H
O
COOH
HO
H H
3
C
O
HO
OH
PRAVASTATIN (PRAVACHOL)
N
OH
CO
2
Na
HO
F
CH( CH
3
)
2
FLUVASTATIN
N
H
3
CO
(H
3
C)
2
HC CH( CH
3
)
2
F
CO
2
Na
OH OH
CERIVASTATIN
Synthetic statins
HMG CoA reductase inhibitors
Precaution:
mild elevation of serum aminotransferase (should
be measured at 2 to 4 month intervals)
minor increases in creatine kinase (myopathy,
muscle pain and tenderness)
do not give during pregnancy
Selected hypolipidemic products
FIBRIC ACID DERIVATIVES
CH
3
H
3
C
O (CH
2
)
3
C
CH
3
CH
3
COOH
GEMFIBROSIL (LOPID)
Cl O C
CH
3
CH
3
COOEt
CLOFIBRATE(ATROMID-S)
Cl
O C
CH
3
CH
3
iPrO
2
C
O
FENOFIBRATE (TRICOR)
Clofibrate (Atromid-S)
Precautions
enhances coumarin activity
renal/hepatic injury contraindication
pregnancy/nursing
cholelithiasis
most commonly reported ADR are GI related
liver malignancies (not very common; but has led to
scant usage)
CLOFIBRATE
Primary activity on triglycerides
MOA:
increases lipoprotein lipase
lowers VLDL
increases peroxisomal FFA oxidation
inhibits cholesterol biosynthesis
increases biliary secretion of cholesterol
ancillary:
decreases platelet adhesiveness/fibrinogen
Gemfibrosil (Lopid)
MOA
stimulates lipoprotein lipase
interact with PPARa (peroxisome proliferator-activated
receptors)
inhibits triglyceride lipolysis in adipose tissue
decreases FFA uptake by the liver
decreases hepatic VLDL/TG synthesis
slight cholesterol lowering effect
precautions
similar to clofibrate
myositis (voluntary muscle inflammation)
GI (indigestion, abdominal pain, diarrhea)
cholelithiasis (increased cholesterol biliary secretion)
half life: 1.1 hours
Fenofibrate (Tricor)
a relatively new fibric acid derivative
(micronized form of the drug)
lowers plasma TG
inhibits TG synthesis
stimulates catabolism of VLDL
indicated primarily for hypertriglyceridemia
same side effects and precaution as in other
fibric acid compounds
half-life: 20 hours
Dose: 67-201 mg/day with meals
Now also available as a 200 mg tablet
NICOTINIC ACID (Niacin)
N
COOH
NICOTINIC ACID (NIACIN)
A water soluble vitamin of the B family;
nicotinamide is not active
Once converted to the amide, it is
incorporated into NAD

In order to be effective, it has to be dosed at the rate of 1.5 to 3.5
gm daily.
A sustained release dosage form is available

adverse effects: GI disturbances (erosion and ulceration)
red flush especially in the face and neck area
caused by vasodilation of capillaries
Nicotinic acid (Niacin)
MOA
dual plasma triglyceride and cholesterol lowering
decreases VLDL and LDL
decreases TG lipase in adipose tissue
increases lipoprotein lipase in adipose tissue
precaution
transient cutaneous flush
histamine release
potentiates BP effect of antihypertensives
Advicor

niacin-extended-release and lovastatin tablets
reduces LDL-C, TC, TG and increases HDL-C
available as 500/20, 750/20 and 100/20 mg
tablets
Rosuvastatin (Crestor)
New statins: rosuvastatin (ZD4522)
N
iPr
COOH
H
S
CH
3
O O
OH
OH
F
nicknamed superstastin/ gorilla statin because of its powerful
effect on LDL cholesterol
Ezetimibe (Zetia)
N
OH
O
F
OH
F
EZETIMIBE
This drug blocks the intestinal absorption of cholesterol. A dose of 10 mg qd leads to
a 19% reduction of LDL; shows real promise in combo product with statins (Schering-
Plough and Merck)
Investigational drugs
acylCoA: cholesterol acyltransferase
inhibitors
Orphan nuclear receptors:
LXR oxycholesterol receptor --- enhanced
cholesterol efflux
FXR bile acid receptor ---- decreased
cholesterol conversion to bile salts
ACAT Inhibitors
S
N
CF
3
O O
CF
3
CF
3
T0901317 -- LXR agonist
CO
2
H
OH
LG268 -- RXR agonist
ACAT Inhibitors
O
CH
CH
3
H
3
C
CH
CH
3
CH
3
S
N
C
H
2
C
O
H
O
CH
CH CH
H
3
C CH
3
CH
3
CH
3
CH
3
H
3
C
O
AVASEMIBE (CI-1011)
Squalene synthase inhibitors
squalestin 1, a fermentation product
derived from Phloma species
(Coelomycetes)
a potent inhibitor of squalene synthase
produces a marked decrease in serum
cholesterol and apoB levels
may represent an alternative clinical
therapy to hypercholesterolemia