Professional Documents
Culture Documents
Lipoprotein metabolism
By
Henry Wormser, Ph.D.
Professor of Medicinal Chemistry
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 Density Diameter Protein % Phospholi Triacylglycerol
class (g/mL) (nm) of dry wt pid % % of dry wt
HDL 1.063-1.21 5 – 15 33 29 8
LDL 1.019 – 18 – 28 25 21 4
1.063
IDL 1.006-1.019 25 - 50 18 22 31
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
Apoproteins of human
lipoproteins
• 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 β-2-
glycoprotein I (involved in TG metabolism)
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
Chylomicron metabolism
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
VLDL metabolism
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 HDL3
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 HDL3 to HDL2a (an HDL
with a high apoE and cholesterol ester content)
• cholesterol ester-rich HDL2a is then converted to
triglyceride-rich HDL2b by concomitant transfer of HDL
cholesterol esters to VLDL and VLDL triglycerides to
HDL
HDL metabolism
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
rate-limiting step
and step subject to
inhibition by statins
Biosynthesis of cholesterol
-synthesis of isopentenyl
-pyrophosphate
A monoterpene
Synthesis of farnesyl
pyroposphate
Biosynthesis of cholesterol
- synthesis of squalene
a sesquiterpene
a triterpene
Synthesis of squalene
Biosynthesis of cholesterol
- synthesis of lanosterol
The demethylation
of lanosterol is also
a useful step for drug
design – i.e. azole antifungals
HO 7a-hydroxylase HO OH
cholesterol 7a-hydroxycholesterol
12a-hydroxylase
O2; NADPH + H+ O2
2 CoA-SH NADPH + H+
2 CoA-SH
OH
C S CoA
C S CoA
O
HO OH
H
cholyl-CoA
HO OH
H
chenodeoxycholyl- CoA
Conversion of cholyl-CoA to
glycocholic acid
ΟΗ
C S CoA
O
glycine ΟΗ
CoA-SH
C N CH2 COOH
HO O
ΟΗ
H
cholyl-CoA
HO ΟΗ
H
glycocholic acid
ΟΗ
Conversion of
C S CoA
cholyl CoA to
O taurocholic acid
HO ΟΗ
H
cholyl-CoA
taurine
CoA-SH
ΟΗ H
HO ΟΗ
H
COO COO
H3N C H + O2 + O2 - CO2
H3N C H H3N CH2
CH2 CH2 CH2
SH SO2 SO3
cysteine cysteine sulfinate taurine
C S CoA
HO OH
H deconjugation +
chenodeoxycholyl- CoA 7a-dehydrxylation
(catalyzed by microbial
enzymes)
COOH
HO
H
C N CH2 COOH
HO ΟΗ
H
glycocholic acid (primary bile acid)
deconjugation +
7α -dehydroxylation
Conversion of glycocholic
( catalyzed by microbial
acid to deoxycholic acid enzymes)
ΟΗ
COOH
• 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
BILE ACIDS
CH3
CH3
HO
CH3
CH3
COOH
COOH
12 CH3 H
CH3 H
H H
H H
3 7
HO OH
H
H
CHOLIC ACID
CHOLANIC ACID
CH3
CH3 CH3
COOH
CH3
COOH CH3 H
CH3 H
H H
H H HO OH
H
HO OH
H
CHENODEOXYCHOLIC ACID (CHENODIOL) URSODEOXYCHOLIC ACID (URSODIOL)
(ACTIGALL)
(CHENIX)
GALLSTONE THERAPEUTIC AGENTS
Type V:
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
HN HN
HN . n HCl . n HCl . n HCl
H H2 H H2N
C C C
. n HCl (CH2)6N(CH3)3 (CH2)9-CH3
OH
H2
C CH
H2C N(CH3)3
(CH2)6N(CH3)3 (CH2)9CH3
n . n HCl
H2N . n HCl . n HCl
HN . n HCl
HN
HN
CHOLESTYRAMINE
COLESEVELAM
Bile sequestering resins
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 • methyldopa
level) • nicotinic acid
• aspirin
• penicillin G
• clindamycin
• propranolol
• clofibrate
• • tetracycline
furosemide
• glipzide • thiazide diuretics
• tolbutamide • digoxin
• phenytoin • hydrocortisone
• imipramine • phosphate supplements
PLANT STEROLS
CH3
CH3
H 3C
H 3C
CH3 CH3
CH3
CH3
CH2CH3 CH2CH3
CH3 CH3
HO HO
HO HO
STIGMASTEROL CAMPESTROL
CH3
H3C
CH3
CH3
CH3
CH3
HO
More plant sterols
CH3
ALPHA1-SITOSTEROL
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
2 NADPH 2 NADP
CH3 CH3
OH OH
COOH COOH
O - CoASH OH
H H
SCoA
NADPH NADPH
CH3 CH3
OH
COOH OH
COOH
OH O
H
SCoA H
O HO O
O O
O CH3
H
H3C H H
CH3
H3C
CH3
H
H3C
Lovastatin
CH3
-10
Ki = 10
HO O
HO
COOH
O
O
H
P
O-
F
HO Cl
N
Et
Cl
Ph
Ki = 10-10 Ki = 10 -11
HO O
HO O
O
O O
O
O
H3C H H O
CH3 H3C H H
CH3
H3C
MEVASTATIN
LOVASTATIN (MEVACOR)
HO O
HO
COOH
O
OH
O
O
O
O
H3C CH3 H
H3C H H
CH3
CH3
HO
SIMVASTATIN (ZOCOR)
PRAVASTATIN (PRAVACHOL)
Synthetic statins
HO
CO2Na
F
OH
OH OH
F
CO2Na
CH(CH3)2
N
H3CO
(H3C)2HC N CH(CH3)2
CERIVASTATIN
FLUVASTATIN
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
CH3
CH3 CH3
O (CH2)3 C COOH Cl O C COOEt
CH3 CH3
H3C
GEMFIBROSIL (LOPID) CLOFIBRATE (ATROMID-S)
Cl
CH3
iPrO2C C O
CH3 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 PPARα (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)
COOH
A water soluble vitamin of the B family;
nicotinamide is not active
N
Once converted to the amide, it is
NICOTINIC ACID (NIACIN)
incorporated into NAD
CH3
F
O S O
N
H
COOH
iPr
OH OH
N
F
O
EZETIMIBE
O O CF3
S
N
CF3
CO2H
T0901317 -- LXR agonist
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