Vitamins

PART II
 Fat soluble vitamins

• are vitamin A, E and K. Vitamin D is considered to be a hormone.

General characteristics:
• apolar hydrophobic molecules,
• are all isoprene [CH2=CH-C(CH3)=CH2 ] derivatives
• require normal fat absorption
• must be transported in the blood, like any other apolar lipid, in lipoproteins or specific binding
proteins.
• can be deposited in small amounts in the body, especially in the liver A, D, K), adipose tissue (E),
membrane level
• suffer specific transformation at the tissue level in order to be eliminated.
• Certain are excreted in the bile, follows the enterohepatic circuit and are excretes in the feces.
• Some metabolites can pass in the urine
 Vitamin A (Retinol , retinoids)

Sources
• Provitamins A (carotenes) are synthesized only by the plants, in the vegetal
products colored in dark green, yellow, red (tomatoes, carrots, peppers).

• retinoides are found only in animal products (liver, fish oils, butter, milk, yolk egg).

• Humans are able to transform carotenes into vitamins A

Vitamin A

Chemical structure of provitamins A

• are yellow to red pigments, with 40 carbon atoms.
• They posses a β-ionone ring and a polyunsaturated side chain which is
composed of isoprenoid units.
H3C
CH3 C 3
H3C CH3 15

β - carotene 6
15' H3C CH3
5 CH C H3
CH3
H3C
CH3 CH3 4'
H3C CH3 15 5'

α - carotene 15' H3C CH3

CH3 CH3
CH3

α - ionone
H3C
CH3 CH3
H3C CH3 15

γ - carotene 15' H3C CH3

CH3 CH3
CH3

β - ionone γ-ionone
Vitamin A

Chemical structure of Vitamins A - retinoids

• 3 biologically active molecules, retinol, retinal (retinaldehyde) and retinoic
acid.
• derived from the plant precursor molecule, β-carotene
• Many different geometric isomers are possible as a result of either a trans
or cis configuration of the double bonds found in the polyene chain.

CH2 OH
7 11
5 CH2 OH
6 9 15 4
13
4 8 10 12 14 3
3 1
2

Vitamin A1 (retinol) Vitamin A2 (3-hydroxyretinol)

CH2
CH2 OH

Vitamin A3
Vitamin A

COOH

Retinoic acid

CH O

11-trans-retinal

O HC

11-cis-retinal
Vitamin A

15 14'
R R β - caroten
14 15'

O2 carotinază

CH O
R
+ R
retinal O HC

NADH + H+ retinal
retinen reductază
(alcool dehidrogenază)

NAD+
R
CH2 OH HOOC
R
retinol acid retinoic
Vitamin A

Metabolism
• Ingested β-carotene is cleaved to yield retinal.
• Retinal is reduced to retinol within the intestines.
• Retinol is esterified to palmitic acid and delivered to the blood via
chylomicrons and uptake by the liver (storage as a lipid ester within
lipocytes).
• Transport of retinol from the liver to extrahepatic tissues occurs by binding
to aporetinol binding protein (RBP).
• Within extrahepatic tissues retinol is bound to cellular retinol binding
protein (CRBP).
• Plasma transport of retinoic acid is accomplished by binding to albumin.
• Some conjugations take place with glucuronic acid, which ensures their
solubilisation and elimination in the urine.
• Unabsorbed carotenoids, as some of degraded products are eliminated in
the feces.
Vitamin A
Biological and physiological role

A. Gene control exerted by retinol and retinoic acid

• Within cells, both retinol and retinoic acid bind to specific receptor proteins.
• the receptor-vitamin complex interacts with specific sequences in several genes involved in
growth and differentiation and affects expression of these genes.
• ⇒ hormones of the steroid/thyroid hormone superfamily.
• ⇒ retinoic acid is involved in the earliest processes of embryogenesis including the
differentiation of the three germ layers, organogenesis and limb development.
B. Additional role of retinol
• in the synthesis of certain glycoproteins and mucopolysaccharides necessary for mucous
production and normal growth regulation.
• This is accomplished by phosphorylation of retinol to retinyl phosphate, which then functions
similarly to dolichol phosphate.
• Retinol and/or retinoic acid are necessary to prevent the synthesis of some high molecular
weight keratins.
• The absence of mucous secretion (produced by certain epithelial cells) has as consequence the
excessive drying of these cells, and the excess of keratin forms keratinized surfaces (hoof
aspect).
• Retinol and/or retinoic acid are necessary for the synthesis of transferrin, the Fe transport
protein.
• The deficiency in these vitamins produce anemia, caused by the faulty mobilization of Fe from
the liver.
Vitamin A

C. Role of Vitamin A in vision

• Photoreception in the eye is the function of two specialized cell types

located in the retina; the rod and cone cells.
• These cells contain a photoreceptor pigment in their membranes.
• The photosensitive compound of most mammalian eyes is a protein termed
opsin to which is covalently coupled an retinal.
• The opsin of rod cells is termed scotopsin.
• The photoreceptor of rod cells is specifically termed rhodopsin or visual
purple - a complex between scotopsin and the 11-cis-retinal (also termed
11-cis-retinene) form of vitamin A.
• Rhodopsin is a serpentine receptor imbedded in the membrane of the rod
cell.
• Intracellularly, rhodopsin is coupled to a specific G-protein termed
transducin.
• Absorption of light ⇒ 11-cis retinal → 11-trans retinal + change in opsin
conformation → cGMP → Na and Ca channels → nerve impuls
Vitamin A
Clinical significances of vitamin A deficiency

• is stored in the liver and deficiency of the vitamin occurs only after
prolonged lack of dietary intake.
• The earliest symptoms :
• night blindness follicular hyperkeratinosis,
• increased susceptibility to infection
• cancer and anemia equivalent to iron deficient anemia (β-caroten –
antioxidant).
• Prolonged lack :
• deterioration of the eye tissue through progressive keratinization of the cornea
- xerophthalmia.
• abnormal formation of keratin in mucous membranes
• failure of bone remodeling, leading to thick, solid bones in the skull with an
increase in cerebrospinal fluid pressure.
• Gonadal dysfunction occurs in males and miscarriage in females.
• Deprivation of vitamin A ultimately results in death.
xeroderma

xerophthalmia
Vitamina A

Excess of vitamin A is toxic.

• limited capacity to metabolize vitamin A,
• excessive intakes lead to accumulation beyond the capacity of binding
proteins, so that unbound vitamin A causes tissue damage.
• Symptoms of toxicity affect :
• the central nervous system (headache, nausea, ataxia, and anorexia, all
associated with increased cerebrospinal fluid pressure),
• the liver (hepatomegaly with histologic changes and hyperlipidemia),
• calcium homeostasis (thickening of the long bones, hypercalcemia and
calcification of soft tissues),
• the skin (excessive dryness, desquamation, and alopecia).
 Vitamin D - calciferols

• Vitamin D is not strictly a vitamin since it can be synthesized in the skin, and under most
conditions that is its major source.
• Only when sunlight is inadequate is a dietary source required.
• Vitamins D are considered to be prehormones D. Also, cholecalciferol (vitamin D3) can be
synthesized by the human body starting from cholesterol.

Sources
• Vegetals contain small amounts of vitamin D, but a high content in provitamin D2 - ergosterol (ex.
yeast).

• Animal food (liver, fish oil, yolk, milk, butter) contain high amounts of vitamin D an also
provitamin D3 - 7,8-dehydrocholesterol.
Vitamin D

Chemical structure of provitamins D

• sterols with 19 carbon atoms in the tetracyclic structure
• a hydroxyl group in position 3,
• two conjugated double bonds in positions 5 and 7 and
• a variable radical in position 17

3HC 3HC
R
3HC 3HC

3HC
17 3HC 3HC
C D CH3
A B
3 5 7
HO HO HO
Ergosterol 7 - dehidrocolesterol
(pro - D2)
(pro - D3)
 Vitamin D
Chemical structure
Chemical structureof Vitamins D D
of Vitamins
• opening
openingofof
ring B and
ring rotation
B and of ring
rotation A with
of ring 180°.180°.
A with
• Vitamin
VitaminD2D2 is ergocalciferol
is ergocalciferol – Vitamin
– Vitamin D3 isD3 is cholecalciferol
cholecalciferol
3HC 3HC

3HC 3HC

3HC 2HC
CH3 CH3
UV

HO HO
Ergosterol Ergocalciferol
(pro - D2) (Vitamina D2)

21 22 3HC 3HC
23
18 20
26
cholesterin– 3HC 3HC

12 17 24 25 dehydrogenase
19 11 13 D 16 3HC 2HC
C
14 15 27 UV
1 9
2 10 8
A B
3 5 7
HO
4 HO HO
6 7 - dehidrocolesterol Colecalciferol
(pro - D3) (Vitamina D3)
cholesterol
 Vitamin D

Metabolism
• The biologically active form - 1,25-dihydroxy
vitamin D3 (1,25-(OH)2D3, also termed D3-25-hydroxylase
calcitriol).
• absorbed at the intestine level (D3 better than
D2).
D3-1α-hydroxylase
• circulate in the blood bound to a α2-globulin.
• Vitamins D are deposited in limited amounts in
liver, kidney, pulmons, brain, teguments (also
small amounts of provitamin D)
Vitamin D

Biochemical and physiological role

• Calcitriol functions in concert with parathyroid hormone (PTH) and

calcitonin to regulate serum calcium and phosphorous levels.
• calcitriol functions as a steroid hormone in inducing the expression of
calbindin, a protein involved in intestinal calcium absorption. Counter ion is
phosphate.
• inhibit calcium and phosphate excretion by stimulating reabsorption by the
distal tubules (kidney)
• ⇒ increases calcemia
• ⇒ sends calcium to bones
• prevents colon cancer
Vitamin D

Clinical significance of vitamin D deficiency

• The main symptom in children is rickets and in adults is osteomalacia.
• Rickets is characterized improper mineralization during the development of
the bones resulting in soft bones.
• Osteomalacia is characterized by demineralization of previously formed
bone leading to increased softness and susceptibility to fracture.
Vitamin D

Vitamin D is toxic in excess

• Some infants are sensitive to intakes of vitamin D as low as 50 μg/day,
resulting in an elevated plasma concentration of calcium.
• This can lead to contraction of blood vessels, high blood pressure, and
calcinosis - the calcification of soft tissues.
• To children, vitamin D excess manifests with sickness, exaggerated thirst,
polyuria, than renal deficiency.
• Although excess dietary vitamin D is toxic, excessive exposure to sunlight
does not lead to vitamin D poisoning because there is a limited capacity to
form the precursor 7-dehydrocholesterol and to take up cholecalciferol
from the skin.
Vitamin E - tocopherols

• also termed the fertility vitamin because is necessary for the reproduction
function of some mammals.
Sources
• include vegetable and seed oils – corn, soy (not fish oil).
Chemical structure
• Tocopherols are methylated derivatives of tocol (6-hydroxy-2-methyl-2-
phytylchroman)
HO
5 4
6 3
CH3
7 1 2
8
O

6–hydroxyl-chroman phytyl
 Vitamin E

• There are several naturally occurring tocophenols (7). All are isoprenoid
substituted 6-hydroxychromanes or tocols.
•
Denomination Structure Relative activity
α- tocopherol 5,7,8-trimethyl-tocol 100

β- tocopherol 5,8-diimethyl-tocol 40

γ- tocopherol 7,8-diimethyl-tocol 8

δ- tocopherol 8-methyl-tocol 1

α-Tocopherol
Vitamin E
Vitamin E

Metabolism

• Bile is required for absorption.

• Approximate 60% of ingested tocopherols are excreted in the feces.
• Vitamin E is absorbed from the intestines packaged in chylomicrons.
• It is delivered to the tissues via chylomicron transport and then to the liver
through chylomicron remnant uptake.
• The liver can export vitamin E in VLDLs.
• Due to its lipophilic nature, vitamin E accumulates in cellular membranes,
fat deposits and other circulating lipoproteins.
• The major site of vitamin E storage is in adipose tissue.
Vitamin E

Biochemical and physiological functions

• The α-tocopherol molecule is the most potent of the tocopherols.
• The major function of vitamin E is to act as a natural antioxidant by
scavenging free radicals and molecular oxygen
CH3

HO
ROOH + Toc - O
ROO + CH3

H3C O R

CH3 CH3
Forma redusã
Reduced formaofvitaminei
Vitamin EE O

ROO + Toc - O ROOH + CH3

H3C O H R

CH3
Forma oxidatã
Oxidized a vitaminei
form of Vitamin EE
Vitamin E
• is important for preventing peroxidation of polyunsaturated membrane fatty acids.
• Active α-tocopherol can be regenerated by interaction with vitamin C following
scavenge of a peroxy free radical.
• Alternatively, α-tocopherol can scavenge two peroxy free radicals and then be
conjugated to glucuronate for excretion in the bile.
• Unlike some other vitamins (niacin, B12, folate) tocopherol is not recycled after
carrying out its function but must be replaced totally to continue its biologic role in
the cell.
• The antioxidant action of vitamin E is effective to high oxygen concentrations, it
tends to be concentrated in those lipid structures that are exposed to the highest O2
partial pressures, eg. the erythrocyte membrane and the membranes of the
respiratory tree.
• Vitamin E and selenium act synergistically and reduce the body’s requirement for
each other. Glutathione peroxidase (selenium) provides a second line of defense
against peroxides before they can propagate in chain reactions, damaging
membranes and other cell components.
• In addition, selenium is required for normal pancreatic function, which is necessary
for the digestion and absorption of lipids, including vitamin E.
• Conversely, vitamin E reduces selenium requirements by preventing loss of
selenium from the body or maintaining it in its active form.
Vitamin E

Clinical significances of vitamin E deficiency

• The major symptom: increase in red blood cell fragility; alteration
phenomena of cardiac and skeletal muscle, with increased creatinine
elimination and installation of muscle dystrophy.
• Other symptoms are neurovegetative troubles, edema, hemolytic anemia,
dysfunctions of the liver, ataxia with retina degeneration.
• Neurological disorders
• Increased intake of vitamin E is recommended in premature infants fed
formulas that are low in the vitamin as well as in persons consuming a diet
high in polyunsaturated fatty acids.

High concentrations of vitamin E

• may have pro-oxidant actions.
• can lead to ovary involution, cycle troubles, nerve affections and to men to
azoospermia.
Vitamin K
Sources
• green vegetables;
• Intestinal bacteria synthesize this vitamin. Vitamin K3 (menadione)
• K1 (phylloquinone) in green vegetables
• K2 (menaquinone) produced by intestinal bacteria;
• K3 is synthetic menadione, menadiol, menadiol acetate
Structure: quinone with isoprenoid side chain

Vitamin K1 Vitamin K2
"n" can be 6, 7 or 9 isoprenoid groups
 Vitamin K
Metabolism
• Bile is required for absorption.
• It is transported with lipoproteins.
• Menadione, being water soluble, is absorbed even in the absence of bile
salts, passing directly into the hepatic vein. Vitamin K3 is alkylated to one of
the vitamin K2 forms of menaquinone at the hepatic tissue level.
• Although vitamin K accumulates initially in the liver, its hepatic
concentration declines rapidly and storage is limited.

Biochemical and physiological functions

• The major function - maintenance of normal levels of the blood clotting
proteins, factors II, VII, IX, X and protein C and protein S, which are
synthesized in the liver as inactive precursor proteins.
• Conversion from inactive to active clotting factor requires a
posttranslational modification of specific glutamate residues – gamma
carboxylation
• Ex. Upon chelation of calcium, prothrombin interacts with phospholipids in
membranes and is proteolysed to thrombin through the action of activated
factor X (Xa).
 Ca2+
Vitamin K COOH O O
γ O C C O
CH2
CH
CH2
CH2
CH
CH
HN CO
HN CO
Glu
Peptidic chain O2 Ca2+ γ γ-carboxyl-Glu
- carboxil - Glu (Gla)
(Gla)
(in lant peptidic)

OH O
COO-
CH3
CH3
O

R R
OH glutamil
Glutamyl- O
carboxilaza
carboxylase Phyloquinone epoxide
filochinona epoxid
CO2

OH O
CH3 CH3

R R
OH O
Vitamin K
OH OH
OH
CH2 CH2

CH2 CO CH3
O O O O O O

Dicumarol Warfarin

The regeneration of the hydroquinone form requires an uncharacterized

reductase. This latter reaction is the site of action of the dicumarol based
anticoagulants such as warfarin.

Vitamin K is involved also in calcium metabolism, in carboxylation of

certain kind of proteins that controls calcium (ex. in bones: osteocalcin and
bone matrix Gla protein. Need also Vit C and Vit D.

Treatment of pregnant women with warfarin can lead to fetal bone

abnormalities (fetal warfarin syndrome).
 Vitamin K

Clinical significance of vitamin K deficiency

• Absorbtion from the intestines only in the presence of bile salts and other lipids through
interaction with chylomicrons.
• ⇒ fat malabsorptive diseases can result in vitamin K deficiency.
• The synthetic vitamin K3 is water soluble and absorbed irrespective of the presence of intestinal
lipids and bile.
• Since the vitamin K2 form is synthesized by intestinal bacteria, deficiency of the vitamin in adults
is rare.
• Long term antibiotic treatment can lead to deficiency in adults.
• Symptoms: hemorrhage
• The intestine of newborn infants is sterile, therefore, vitamin K deficiency in infants is possible if
lacking from the early diet. The primary symptom of a deficiency in infants is a hemorrhagic
syndrome.
Cofactors of vitamin type

• act similarly to vitamins,

• human body has the capacity to synthesize them.
• In some cases, due to some defects in their synthesis, their extern intake
becomes absolutely necessary, the compounds becoming real vitamins.
• Some of these compounds are:
• niacin (so termed vitamin B3),
• carnitine,
• ubiquinone,
• lipoic acid,
• tetrahydrobiopterine,
• inositol, etc
 Ubiquinone

• It is synthesize in cytoplasm and microsoms, starting from tyrosine and pharnesylpirophosphate

(intermediary compound in cholesterol synthesis).
• Ubiquinone is free in the internal mitochondrial membrane where it acts as electron transporter
between complex I and complex III, in the respiratory chain.
• It participates, in the cytoplasm, to the control of redox processes.

O
H3C O CH3
n = 10 ⇒ coenzyme
Q10 in humans
H3C O CH2 CH C CH2 H
O CH3
L- Carnitine
Sources:
• animal products (meat).
• very small amounts of it are found in plants, with few exceptions, such as avocado and
some fermented soy products.
Structure
• Trimethylated, quarternary amine

CH3

Synthesis: OOC
from L-lysine +and needs:
CH2an L-methionine
CH CH2 N CH3 magnesium, iron, ascorbic acid,
niacine, pyridoxal-phosphate, 5-methyl-tetrahydrofolate, methyl-cobalamine and
bethaine
OH CH3
Cardiac and skeletal muscles cannot synthesize carnitine,
Premature and newborn children have a low synthesis, extern intake being necessary.
L-carnithine

Biochemical Role

• Transport of long chain fatty acids through mitochondial

membrane
• Fatty acids are oxidised in mitochondria ⇒ energy
• to remove short-chain and medium-chain fatty acids from the
mitochondria in order to maintain coenzyme A levels in these
organelles.
• Antioxidant properties
• cardioprotective effect - reduces the sanguine level of
triacylglycerols and increases HDL-cholesterol level.
• neuroprotective activity
.
L-carnithine

Deficiency
Causes
• Inborn deficiency in synthesis
• associated with dialysis, although intestinal resection, severe infections,
and liver disease
Effects
• muscle fatigue, cramps, and myoglobinemia following exercise hypo-
glycemia, progressive myasthenia, hypotonia, or lethargy
• accumulation of neutral lipid within skeletal muscle, heart tissue and liver, a
disruption of muscle fibers, and an accumulation of large aggregates of
mitochondria within the skeletal and smooth muscle.
• ⇒ cardiomyopathy, congestive heart failure, encephalopathy,
hepatomegaly, impaired growth and development in infants, and
neuromuscular disorders.
 α-Lipoic acid (thiooctacid)

• Present in all cells

• Chemical structure
• Lipoic acid is the 6,8–dithiooctanoic acid.
• It is a mixture of two structures, one with a closed ring with a disulfide bridge; the other form, the
reduced one, has an open ring:
S S SH SH

H 2C CH (C H 2 )4 C OOH H 2C CH (C H 2 )4 C OOH
CH2 CH2

oxidized form reduced form

Lipoic acid

Biochemical role
• It is covalently attached (an amide bond) to a ε–amine of a lysine residue
which is part of an enzyme structure.
• multienzymatic complex with a role in the decarboxylation of α–ketoacids.
Beside lipoic acid, the enzyme contains subunits with thiamin-
pyrophosphate, CoA, FAD and NAD (see vitamin B1).
• Example: pyruvate dehydrogenase - contains a 4 enzymes

pyruvate + CoA + NAD+ → acetyl–CoA + CO2 + NADH + H+

• α–ketoglutarate dehydrogenaseO - decarboxylation of α–ketoglutarate to

succinyl-CoA (citric acid cycle)
H 3C C
O S
O
SH
H 3C C C O O- T PP CoA–SH
H 3C C ~S CoA
R +
CO2 C H3 S S
SH
E3 SH
CH OH H2C CH R E2

CH2 F AD H 2 FA D
T PP E1 R
Tetrahydrobiopterin- BH4

• Found in all cells of superior animals

Chemical Structure
• contains a reduced pteridinic nucleus with a 1,2-propanediol attached to
position 6.
O H H
NH
HN 5 C C CH3
6
H2N 7 OH OH
N NH
8
tetrahydrobiopterine

Metabolism
• synthesized from guanosine triphosphate (GTP) – needs NADPH.
• catabolism - removing the side chain and desamination.
• The catabolism products are eliminated in urine.
Tetrahydrobiopterin
Biochemical role
• BH4 is one of the strongest reducing natural agents.
• cofactor for:
• phenylalanine-4-hydroxylase (PAH) ⇒ Tyr
• tyrosine-3-hydroxylase ⇒ catecholamines
• tryptophan-5-hydroxylase ⇒ serotonin
• hydroxylation of aromatic amino acids,
• nitric oxide synthase ⇒ NO ⇒ strong vasodilator effect
• glyceryl-ether monooxygenase
• increases the proliferation rate of erythroid cells
• increases the expression of phenylalanine hydroxylase
• Vitamin C increases the BH4 synthesis and stabilizes this compound
(antioxidant protective effect).
• Insulin also activates BH4 synthesis.
• In diabetes mellitus, BH4 improves vasodilatation but also corrects the
increased oxidation of NADPH
Tetrahydrobiopterin

Guanosine triphosphate (GTP)

Neopterin Dihydroneopterin triphosphate

6 - Pyruvoyl - tetrahydropterin

Tyrosine Tryptophan
Phenylalanine ARG
Tetrahydrobiopterin
Tyrosine Triptophan
hydroxylase hydroxylase
Phenylalanine-
hydroxylase
L - DOPA 5 - OH - Tryptophan NO synthase
Dihydrobiopterin Pterin - 4a - carbinolamine
Tyrosine

Dopamine Serotonin Citrulline + NO

Biopterin Primapterin

Catecholamines Neurotransmiţters
 Tetrahydrobiopterin

Deficiency
• appears at low 17β-estradiol plasma concentrations, in hypercholesterolemia, treatment with
glucocorticoids, deoxycorticosterone, dexamethasone.

Effects:
• modification of the ratio superoxyde anion/NO and, as a consequence, an endothelial dysfunction
appears with vasoconstriction and increasing of blood pressure.
• BH4 deficiency is an attractive hypothesis for the cause of impaired vasodilatatory responsiveness
in certain clinical settings such as the insulin-resistant state, diabetes, or atherosclerosis.
• Deficiency in DOPA synthesis is the basis of serious anomalies – DOPA and tyrosine deficit,
accumulation of phenylalanine and its impaired catabolism, manifesting phenylketonuria.