tized photodestruction of an essential histidine. J 15. Vannucchi H, Kutnink M, Sauberlich HE.

Interac-
Biol Chem. 1979;254:6489-95
13. McCormick CC, Menard MP, Cousins RJ. Induc-
tion of hepatic metallothionein by feeding zinc to
rats of depleted zinc status. Am J Physiol 1981;
240zE414-21
14. Grider A, Cousins RJ. Zinc-induced changes in
human erythrocyte metallothionein. FASEB J
1989;3:A1078
tion among niacin, vitamin B, and zinc in rats re-
ceiving ethanol. Int J Vitam Nutr Res 1986;56:
355-62
16. Vannucchi H, Moreno FS. Interaction of niacin
and zinc metabolism in patients with alcoholic
pellagra. Am J Clin Nutr 1989;50:364-9
17. Dunn MA, Blalock TL, Cousins RJ. Metallothi-
onein. Proc SOCExp Biol Med 1987;185:107-19

AMlNOCARNlTlNE AND RELATED COMPOUNDS AS INHIBITORS OF

CARNlTlNE TRANSFERASES: PHYSIOLOGIC IMPLICATIONS
p-Aminocarnitine and its N-acetyl and N-palmitoyl amides were examined as
inhibitors of carnitine acetyltransferase, carnitine palmitoyltransferase,
and of fatty-acid oxidation in whole animals, tissues, and hepatic
microsomal systems. Results were consistent with subsequent
findings that aminocarnitine and palmitoylcarnitine have
significant antiketogenic and hypoglycemic effects
in experimental animals.

The close interaction between metabolism
of glucose and that of fatty acids in dia-
betes has long been recognized. Thus, in
insulin deficiency, fatty-acid oxidation is
increased, with a concomitant rise in ke-
tone bodies in the blood. Conversely, utili-
zation of glucose is slowed, gluconeogen-
esis is accelerated, and hyperglycemia
develops. It is not surprising that consider-
able research has been directed to discov-
ery of compounds that might inhibit the ox-
idation of free fatty acids (which are
abnormally increased in concentration)
and that would have some application in
the treatment of diabetes. In this context,
recent research from laboratories in
Japan'f2 and the United States3r4 that has
led to the independent discovery and prop-
erties of p-amino analogs of carnitine is of
particular interest.
In the Japanese study,' a bioautographic
screening procedure (similar to antibiotic
screening techniques) was devised in a
search for metabolic inhibitors of fatty-acid
oxidation in microorganisms. The yeast
Candida albicans was seeded heavily in
minimal agar medium at 45°C; the medium
contained either oleic acid or glucose as
the sole source of carbon. After cooling
and solidification of these media, paper
chromatograms of microbial extracts were
placed on the agar plates. After incubation
only zones of inhibition observed on oleic
acid-containing plates were considered.
Such zones of inhibition should have de-
rived from microbial products interfering
specifically with oleate catabolism (oxida-
tion), a process essential for growth of C.
albicans. By this technique it was found
that Emericella quadrilineata produced an
inhibitor termed emericedin, which has the
structure (R)-3-acetylamino-4-(trirnethylam-
monio)-butyric acid. Hydrolysis of emerice-
din gave the free base emeriarnine.
CH3
I
CH3- N+ - CH2- CH - CHZ- COO-
I I
CH3 NH
I
R
(Emericedin: R = -COCH3; Emeriamine: R
= -H)

258 NUTRITION REVI€WS/VOL 48. NO GIJUNE 1990

 The close chemical relation of these
compounds to carnitine and acetylcarni-
tine prompted detailed studies of their ef-
fect in carnitine acyl transfer systems. The
p-N-palmitoylamide of emeriamine, pal-
mitoylemeriamine, obtained via chemical
synthesis, was also included in this study.
Acyl-SCoA + Carnitine e Acyl Carnitine +
CoASH
(Carnit ineacetyItransferase5 (CAT): Acy I =
Acetyl ; Carnitine palm itoyltransferase6
(CarPamTase): Acyl = Palmitoyl)
Emeriamine strongly inhibited oxidation
of fatty acids in rat liver homogenates but
was less effective in heart and muscle ho-
mogenates. In rat liver mitochondria 3.2 p~
emeriamine gave 50% inhibition of carni-
tine-dependent [1-I4C] palmitate oxidation
to 14C02but had no effect on oxidation of
[1-14C] octanoate; this result suggests ex-
clusive interference with oxidation of long-
chain fatty acids by CarPamTase. In this re-
gard, 62.5 p , ~emeriamine gave 50%
inhibition of CarPamTase I. An interesting
facet of this work1.* was the finding that al-
though emeriamine itself did not inhibit
CAT, emericedin did effectively inhibit CAT.
Indeed, Kanamaru and Okazakil showed
that hepatic mitochondria (as a source of
CAT and CarPamTase), when incubated
under appropriate conditions with emeri-
amine, formed emericedin and palmitoyl-
emeriamine, which were potent inhibitors
of their respective carnitine transferases.
Thus, emeriamine appears to be a suicide
substrate, which, after appropriate incuba-
tion in the CAT or CarPamTase system,
produces products that inhibit these re-
spective transferases.
In an entirely independent study, Jenkins
and Griffith3 synthesized a-aminocarnitine
and acetyl-DL-aminocarnitine because they
expected that the latter carnitine analog, in
contrast to acetylcarnitine, would be ther-
modynamically and metabolically stable
and might have some utility as an enzyme
or transport inhibitor in relevant carnitine
transferase systems. In many respects, the
effects of these synthetic analogs on CAT
and CarPamTase confirmed and comple-
mented the observations of the Japanese
group,’S2 and only certain results of the
American workers4 will be discussed. They
found that acetyl-DL-aminocarnitine was a
potent competitive inhibitor of CAT (Ki =
24 p , ~ ) which
, bound to the enzyme about
13-fold more tightly than did acetylcarni-
tine. These workers also synthesized pal-
mitoyl-m-aminocarnitine and showed that
it was about 100-fold more effective than
m-aminocarnitine as an inhibitor of Car-
PamTase.
Another important finding of Jenkins and
Griffith4 was that small doses of aminocar-
nitine and palmitoylaminocarnitine pre-
vented the development of ketoacidemia in
fasted normal mice and reversed the keto-
acidemia observed in fasted diabetic mice.
Aminocarnitine had a strong hypoglycemic
effect in fasted diabetic mice, e.g., a single
dose (0.3 mmol/kg) normalized plasma glu-
cose levels within 4-8 h and remained ef-
fective for a12 h. Similar results were
found by the Japanese investigator^,^^^^^
who evaluated emeriamine in rats with hy-
poglycemia and ketonemia. In these stud-
ies the hypoglycemic and antiketogenic ef-
fects of emeriamine, at a dose of 10 mg/kg
(0.06 mmollkg), lasted for >6 h.
CarPamTase, as the rate-limiting step of
fatty-acid oxidation, has served as a focal
point for many investigators in the develop-
ment of inhibitors to manipulate the activity
of this key enzyme. The potential therapeu-
tic use of certain of these compounds in
treatment of diabetes mellitus has been re-
viewed.* The research discussed herein
clearly shows that aminocarnitine and pal-
mitoylcarnitine have significant antiketo-
genic and hypoglycemic effects in experi-
mental animals. Perhaps a major limitation
in the use of these drugs in diabetes is the
observation of Jenkins and Griffith4 that
these aminocynitines inhibit the in-vivo
oxidation of palmitate to COP by only
-50%, implying substantial non-carnitine-
mediated oxidation of fatty acids, e.g., via
substantial peroxisomal oxidation. More-
over, as might be anticipated, the effective
NUTRITION REVIEWSIVOL 48, NO GIJUNE 1990 259
 inhibition of CarPamTase by DL-aminocar-
nitine and palmitoyl-DL-aminocarnitine, at
levels where significant antiketogenic and
hypoglycemic effects were seen (fasted
mice), caused marked hepatic elevation of
triacylglycerols. Such findings add a note
of concern to this approach to diabetes
therapy. In this regard, the suggestiong that
CarPamTase inhibitors be used synergisti-
cally with insulin seems logical and worthy
of investigation.
It is interesting to reflect on the produc-
tion of emericedin by Emericella quadrili-
neata. What is the purpose of such diaboli-
cal biosynthesis in nature? Whatever the
answer, this field of research concerning
the fashioning of CAT and CarPamTase in-
hibitors, by either humans or microbes, is
an attractive field for further investigation
because it has ramifications both for basic
science and clinical practice.
1. Kanamaru T, Okazaki H. Emeriamine: a new inhib-
itor of long chain fatty acid oxidation and its anti-
diabetic activity. In: Demain AL, Somkuti GA,
Hunter-Cevera JC, Rossmore HW, eds. Novel mi-
crobial products for medicine and agriculture.
New York: Elsevier, 1989:135-144
VITAMIN C REGULATION OF CARTILAGE MATURATION
Treatment of chondrocytes with vitamin C under defined conditions in cell culture
initiates a differentiation program that closely mimics the normal maturation of
cells in the growth plate of long bones.
In scurvy, a vitamin C-deficiency disease,
fundamental defects are found in connec-
tive tissues including cartilage and bone. It
has been assumed for many years that the
clinical manifestations of scurvy are di-
rectly related to changes in collagen me-
tabolism that accompany ascorbate defi-
ciency. However, little is known about the
cellular changes that accompany the dis-
eased state. Two recent studies that exam-
ined effects of vitamin C on cultured chon-
drocytes indicated that this vitamin induces
260 NUTRITION REVIEWSIVOL 48,NO GIJUNE 1990
2. Kanamaru T, Shinagawa S , Asai M, et al. Emeri-
amine, an antidiabetic P-aminobetaine derived
from a novel fungal metabolite. Life Sci 1985;37:
217-23
3. Jenkins DL, Griffith OW. DL-Aminocarnitine and
acetyl-DL-aminocarnitine. Potent inhibitors of car-
nitine acyltransferases and hepatic triglyceride ca-
tabolism. J Biol Chem 1985;260:14748-55
4. Jenkins DL, Griffith OW. Antiketogenic and hy-
poglycemic effects of aminocarnitine and acyl-
aminocarnitines. Proc Natl Acad Sci USA 1986;83:
290-4
Bieber LL, Lewin LM. Measurement of carnitine
and 0-acylcarnitine. Methods Enzymol 1981;72:
276-87
Bremer J. Carnitine in intermediary metabolism;
the biosynthesis of palmitoylcarnitine by cell sub-
fractions. J Biol Chem 1963;238:2774-9
Harano Y, Kojima H, Kashiwagi A, et al. A new car-
nitine analogue for the correction of metabolic de-
rangements in diabetes. In: Sakamoto N, Alberti
KGMM, Hotta N, eds. Recent trends in manage-
ment of diabetes mellitus. Amsterdam: Excerpta
Medica, 1987:567-71
Tutwiler GF, Kirsch T, Mohrbacher RJ, Ho W.
Pharmacologic profile of methyl P-tetradecylglyci-
date (McN-3716)-an orally effective hypoglyce-
mic agent. Metabolism 1978;27:1539-56
McGarry JD, Foster DW. Acute reversal of experi-
mental diabetic ketoacidosis in the rat with (+)-
decanoylcarnitine. J Clin Invest 1973;52:877-84
complex changes in cell differentiation that
are necessary for cartilage maturation in
vitro and may be important for normal bone
growth.’,*
In growing animals the ends of long
bones contain regions known as epiphy-
seal growth plates where chondrocytes me-
diate longitudinal growth by proliferating
and forming a type II collagen-containing
extracellular matrix. Over time, this matrix
becomes mineralized and is eventually re-
placed with bone, which contains mineral-