SULFONAMIDES AND

DHFR INHIBITORS
VMED5223
Veterinary Pharmacology
10-2007
Masami Yoshimura
myoshimura@vetmed.lsu.edu
Room2423
 History

n 1935 - discovery of the antibacterial nature of

prontosil (prodrug, "bioactivation") by Gerhard
Domagk.
n Late 1930s-early 40s - more than 5,000
compounds were prepared. (Sulfa drugs)
n 1939 - Domagk received the Nobel Prize in
Medicine for his discovery of the first drug effective
against bacterial infections.
n mid-1970s - introduction of the combination of
trimethoprim and sulfamethoxazole has increased
the use of sulfonamides.
Prontosil
 Mechanism of Action
PABA is an essential nutrient of many bacteria
in the synthesis of folic acid. Sulfanilamide
binds to the enzyme that converts PABA to
folic acid. (Antimetabolite)
Structure of Sulfonamides
SULFANILAMIDE SULFAMETHAZINE SULFAQUINOXALINE SULFATHIAZOLE

SULFASALAZINE SULFADIAZINE SULFISOXAZOLE

 Where is selective toxicity?

Folate
(Vitamin B9)
 Folate (Vitamin B9) Deficiency
Adequate levels of folate are particularly critical
during the development of the nervous system. A
shortage of folate in this period can cause neural-tube
defects, including spina bifida.

Folate deficiency is probably the most common

vitamin deficiency. Since addition of folate to flour in
the U.S. began in 1996, neural-tube birth defects have
decreased by ~20%. A recent study concluded that
adding folate to food also has decreased deaths from
heart attacks & stroke by 48,000/year.
 Folic Acid: Vitamin Prevents
Serious Birth Defects
n Neural tube defects (NTDs) are among the
most serious and common birth defects in
the United States. Each year, an estimated
2,500 babies are born with these defects,
and many additional affected pregnancies
result in miscarriage or stillbirth. The most
common NTD is spina bifida, a leading
cause of childhood paralysis.
Information provided by
The March of Dimes
 Folic Acid: Vitamin Prevents
Serious Birth Defects
n Only 32% of non-pregnant women between the
ages of 18 and 45 took a daily multivitamin
containing folic acid.
n 75% of women say they are aware of folic acid.
n 90% do not know that folic acid must be
consumed before pregnancy to be effective.
n 14% know that folic acid prevents birth defects.

Information provided by
The March of Dimes
 Spectrum/Clinical use
n Bacteriostatic
n Combined with DHFR inhibitor (potentiated
sulfonamides) – bactericidal
n Spectum – broad
n Treatment of infections of
n CNS
n respiratory tract
n GI tract
n urinary tract
 Absotption/Distribution
n Well absorbed from GI tract (systemic
sulfas) except enteric sulfa
n Administration
n oral, i.v., topical, mammary infusion
n Widely distributed
n pleural, ocular, synovial, cerebrospinal fluid
n fetal circulation, milk
 Matabolism/Excretion
n Major metabolite = N-acetyl sulfonamide
n Dog cannot
n Less soluble
n Excreted by kidney (parent drugs and
metabolites)
n Enteric sulfas to feces
 Toxicity
n Urinary tract
n crystalluria
n Hematopoietic system
n hypoprothrombinemia, aplastic anemia,
thrombocytopenia
n Hypersinsitivity reaction
n rashes, fever, hepatitis
n Keratoconjunctivitis Sicca (KCS) for dogs
 Microbial Resistance
n Alteration of dihydropteroate synthase
n Increased production of PABA
n Impaired drug penetration into the
microbial cell
 DHFR Inhibitors
n Structural analogues of DHF
n Trimethoprim (diaminopyrimidene derivative)
n Competitive inhibitor of DHFR
n Bacteriostatic
n Combined with sulfonamides (potentiated
sulfonamides) – bactericidal - synergistic
effect
 Folate
(Vitamin B9)
 Selective Toxicity of
Trimethoprim
Source of enzyme Trimethoprim EC50 (nM)

Bacterial
Escherichia coli 5
Staphylococcus aureus 15
Proteus vulgaris 5
Mammalian
Rat 260,000
Rabbit 370,000
Human 300,000
DHFR Inhibitors

DHF

Trimethoprim

Methotrexate
 Toxicity/pharmacokinetics
n nausea and diarrhea
n Absorbed from GI tract
n Well distributed
n Metabolism in liver
n Excreted in urine
 Spectrum/Clinical use
n Spectum – broad
n Used as potentiated sulfonamides
n bactericidal
 Potentiated Sulfonamides

MIC (
g/ml)
Sulfamethoxazole Trimethoprim
Organism alone mixture alone mixture

Staphylococcus aureus 3 0.3 1 0.015

Streptococcus pneumoniae 30 2 2 0.1

Haemophilus influenzae 10 0.3 1 0.015

Shigella sonnei 10 1 0.3 0.05

Proteus vulgaris 30 3 3 0.15

 Residues in Food Animals
n Used as food/drinking water additives
n Potential health hazard to human
n Sulfonamide residues produced more
drug-residue violations than any other
drug.
n Pork, veal, poultry