Method for preventing body fat deposition in mammals

A method for reducing the rate of liver triglyceride synthesis and body fat deposition in mammals by orally administering over a prolonged period a therapeutic mixture of effective amounts of pyruvate and dihydroxyacetone to which may be added riboflavin. The method also has the effect of increasing the glycogen-storing capabilities of the liver and increasing the body protein concentration.

BACKGROUND OF THE INVENTION 
In U.S. Pat. No. 4,158,057, issued June 12, 1979, a method is described for 
preventing the accumulation of excessive fatty deposits in the livers of 
mammals. It has long been known that ingestion of ethyl alcohol in 
mammals, including man, frequently results in the accumulation of 
excessive fatty deposits in the liver. In many cases, this accumulation 
tends to become irreversible and may lead to serious consequences, 
particularly alcohol-induced hepatitis and, ultimately, cirrhosis. 
The invention described in the aforesaid U.S. Pat. No. 4,158,057 resides in 
the discovery that excessive fatty deposits in the liver can be reduced or 
prevented from occurring by administering a therapeutic composition 
consisting of a mixture of pyruvate and dihydroxyacetone to which may be 
added riboflavin. These substances are natural metabolites which occur in 
the body as a result of normal digestive processes. Heretofore, however, 
there has been no appreciation of any correlation between the accumulation 
of fatty deposits in the liver, usually due to the ingestion of alcohol, 
and the accumulation of fat in other parts of the body. 
SUMMARY OF THE INVENTION 
In accordance with the present invention, it has now been found, quite 
surprisingly, that the mixture disclosed in U.S. Pat. No. 4,158,057, when 
administered for a relatively long period of time, at least 15 days or 
more, results in a reduction of the rate of hepatic triglyceride 
generation and body fat deposition for a given diet. The invention is thus 
useful for impeding overweight conditions in mammals, with or without 
ingestion of ethanol. 
Additionally, it has been found that prolonged ingestion of a mixture of 
pyruvate and dihydroxyacetone, with or without riboflavin, increases the 
glycogen-storage capabilities of the liver. Stored glycogen is thus 
increased for subsequent release into the bloodstream. Stored glycogen has 
been reported to increase the performance and endurance of athletes. 
A further surprising discovery of the present invention is that there is a 
decrease in the total body fat with a secondary inhibition of weight gain 
in mammals. Prolonged ingestion of a mixture of pyruvate and 
dihydroxyacetone, with or without riboflavin, actually changes the body 
composition to the extent that body fat is actually decreased by the 
inhibitory effect of the lipotropic agent on fat metabolism. Moreover, a 
significant finding of the present invention is a small but clinically 
substantial increase in body protein concentration induced by the 
lipotropic agent. 
The above and other objects and features of the invention will become 
apparent from the following detailed description taken in connection with 
the accompanying drawings which form a part of this specification, and in 
which:

To demonstrate the efficacy of the invention, a group of rats (Group A) 
each weighing about 200 grams was fed a standard laboratory diet for a 
period of 60 days, the diet containing 15% protein, 28% fat and 57% 
carbohydrate. A second group of rats (Group B) was fed the same diet as 
Group A except with the addition of a mixture of pyruvate, 
dihydroxyacetone and riboflavin. The specific mixture comprised 22.5 grams 
of pyruvate, 22.5 grams of dihydroxyacetone and 2.25 grams of riboflavin 
per 1000 cubic centimeters of diet. After being on the aforesaid diets for 
60 days, each group of rats was injected with radioactive glycerol. About 
one hour after the injection, the rats were sacrificed, their livers 
removed, and the radioactive triglyceride generated was determined by 
chemical analysis. The results are shown in FIG. 1; and it will be noted 
that the rats in Group B which ingested the mixture of pyruvate, 
dihydroxyacetone and riboflavin with the same basic diet had a much lower 
rate of liver triglyceride synthesis. That is, those in Group B had a 
synthesis rate of about 0.05 millimol per gram per 30 minutes; while those 
which did not receive the mixture in Group A had a much higher 
triglyceride synthesis rate of 0.15 millimol per gram per 30 minutes. 
More surprising is the effect on weight gain by adding the mixture of 
pyruvate, dihydroxyacetoned and riboflavin to the diet. This is shown in 
FIG. 2 where, it will be noted, those rats which did not receive the 
mixture gained almost 190 grams during the 60-day period; whereas those 
which did receive the agent (Group B) gained only about 160 grams. From 
this it can be concluded that as the rate of triglyceride synthesis 
decreases, so also does the total weight gain. 
In another group of experiments extending over a period of one year, rats 
were divided into the groups ranging from 4 to 8 in number. The first 
group (Group C) was fed a diet comprising 15% protein, 28% fat and 57% 
carbohydrate, the same as for Groups A and B. The second group (Group D) 
received the same diet as Group C except that half of the carbohydrate 
content of the diet was substituted isocalorically with ethanol. The third 
group (Group E) received the same dies as Group D containing ethanol but 
with the addition of 22.5 grams of pyruvate, 22.5 grams of 
dihydroxyacetone and 2.25 grams of riboflavin per 1000 cubic centimeters 
of diet. The effect of the rate of triglyceride synthesis is shown in FIG. 
3. Note that Group D, which ingested ethanol, had a much higher rate of 
triglyceride synthesis; whereas Group E which ingested ethanol but at the 
same time ingested the treating agent of the invention had a much, much 
lower rate of triglyceride synthesis. The synthesis rate was determined 
with a radioactive precursor in the same manner as described above in 
connection with Groups A and B. The effect on weight gain is shown in FIG. 
4. Note that Groups C and D had the same weight gain over a year's time, 
which indicates that ingestion of ethanol and the rate of triglyceride 
synthesis have very little to do with weight gain. In Group E, however, 
which had the same diet as Group D, weight gain is significantly lower, 
being on the order of 450 grams per year as contrasted with 600 grams per 
year for Groups C and D. 
While the treating agent in all cases contained riboflavin, it is believed 
that this latter agent has a minimal effect on weight gain and that 
substantially the same effect can be obtained with or without the addition 
of riboflavin. The quantitative effect on weight gain is dependent upon 
the dosage; however the dosage is not critical per se. In order to obtain 
any practical effect as regards weight loss, the agent of the invention 
should be administered, usually for at least 15 days, until a perceptible 
weight loss is observed for a given diet. An effective treatment for 
reducing weight gain in mammals is, therefore, provided utilizing natural 
metablities readily available at a relatively low cost. 
As is known, the liver, in addition to synthesizing triglycerides, also 
acts as a storage medium for glycogen. Glycogen is known as the emergency 
fuel since, unlike fat stores, glycogen is readily available and easy to 
convert back into glucose. That is, glucose brought to the liver from the 
intestine via the portal vein is converted to glycogen and stored. As the 
need arises, glucose is re-formed from glycogen and released into the 
bloodstream. It has been found that by administering the mixture of the 
invention over a long period of time, the glycogen-storing capability of 
the liver is increased, accompanied by an increase in the size of the 
liver. This is shown in the following Table where the glycogen 
concentration in mg/g of liver tissue and total glycogen are tabulated for 
the same groups of rats C, D and E described above in connection with FIG. 
2, the rats being treated for a period of one year. The livers of four 
rats in each group were subjected to a standard acid extraction of 
glycogen after sacrifice. 
TABLE 
______________________________________ 
GROUP C GROUP D GROUP E 
______________________________________ 
Glycogen Concentration* 
Rat No. 1 13.2 16.5 37.1 
Rat No. 2 17.9 13.6 31.1 
Rat No. 3 29.9 10.4 80.8 
Rat No. 4 25.6 30.2 40.3 
Average 
value (mg/g) 
21.65 17.6 47.3 
Total Glycogen** 
Rat No. 1 227.7 275.3 841.4 
Rat No. 2 225.5 199.6 602.0 
Rat No. 3 488.8 182.8 1624.8 
Rat No. 4 502.0 750.1 961.9 
Average 
value (mg) 361 351 1007 
______________________________________ 
*mg of glycogen per gram of liver 
**mg of glycogen 
While the effect on individual rats in each group varies substantially, it 
can be seen from the foregoing Table that all rats in Group E which were 
treated with the mixture of the invention without ingestion of ethanol had 
much higher glycogen contents than either those in Group D which ingested 
ethanol without treatment and those in Group C which neither ingested 
ethanol nor were treated with the mixture of the invention. 
For many years athletes have attempted by eating large amounts of 
carbohydrate-laden meals, to increase their glycogen stores prior to an 
athletic event requiring long endurance or sustained high performance. The 
use of the mixture of the invention appears to increase greatly the 
glycogen-storage capability of the liver. Thus, by administering a dosage 
of pyruvate and dihydroxyacetone to people prior to a strenuous vent may 
increase endurance and/or performance. Riboflavin can be added to the 
dosage. 
To further demonstrate the efficacy of the invention, a group of six rats 
(Group F) each weighing about 200 grams was fed a standard laboratory diet 
for a period of 120 days, the diet containing 15% protein, 28% fat and 57% 
carbohydrate. A second group of six rats (Group G) was fed the same diet 
as Group F except with the addition of a mixture of pyruvate, 
dihydroxyacetone and riboflavin. The specific mixture comprised 22.5 grams 
of pyruvate, 22.5 grams of dihydroxyacetone and 2.25 grams of riboflavin 
per 1000 cubic centimeters of diet. After being on the aforesaid diets for 
120 days, the weight gain results are shown in FIG. 5; and it will be 
noted again that the rats in Group G which ingested the mixture of 
pyruvate and dihydroxyacetone and riboflavin with the same basic diet had 
a lower weight gain, i.e., about 495 grams versus about 585 grams for the 
rats in Group F. 
Again, there was found a marked inhibition of weight gain in animals fed 
the agent pyruvate, dihydroxyacetone and riboflavin. Of interest is the 
finding that this weight inhibition that occurs is actually secondary to a 
decrease in total body fat. From FIG. 5, it can be seen that the 
lipotropic agent induces a 16% decrease in weight gain while, as shown in 
FIG. 6, there was a 32% decrease in total body fat in the rats of Group G 
as compared with the rats of Group F. The total body fat in the six rats 
comprising Group F was about 235 grams; whereas the total body fat in the 
rats of Group G was about 160 grams. Actual changes to the body 
composition were found in the rats of Group G, and the percent of body fat 
decreased by about 7%. 
As shown by the bar graph of FIG. 7, the rats of Group F experienced about 
a 31.4% increase in body fat; whereas the rats of Group G which ingested 
the mixture of pyruvate, dihydroxyacetone and riboflavin experienced a 
21.7% increase. From this data, it is concluded that the lipotropic agent 
not only inhibits weight gain but that this decrease in weight gain is 
secondary to an inhibition of body fat; not protein, water, carbohydrate 
or minerals. The inhibitory effect of the lipotropic agent on fat 
metabolism is so great that the body composition actually is changed to 
the extent that body fat percent is decreased. 
Another surprising finding of the present invention is an increase in body 
protein concentration (2%) induced by the lipotropic agent. Although this 
increase is small, it is clinically substantial since the protein 
concentration of the body is small and only a modest increase in body 
protein is helpful. This is especially important in certain patients, 
specifically weight-watchers or body-builders. The body protein of the 
rats in Group F, as shown in FIG. 8, was about 10.5%; whereas the body 
protein of the rats in Group G was about 12.5%. 
In the preferred aspect of the present invention there is provided a 
container or package including a pharmaceutically-acceptable mixture of 
pyruvate and dihydroxyacetone with or without riboflavin in a unit dosage 
quantity together with instructions for administration of effective 
quantities over a period of time, usually at least 15 days. Preferably, 
the container includes a liquid base, such as milk or glucose, to improve 
palpability and/or patient acceptance of the mixture. Depending, of 
course, upon the circumstances, an effective daily dosage may be divided 
in two containers. The contents of the container may be concentrated 
without dispersion in a liquid base to reduce bulk for conveniently 
supplying a sufficient quantity for an effective daily usage over a period 
of at least 15 days. This also offers the advantage that the dosage which 
may vary from patient-to-patient can be conveniently selected. Daily 
dosages may be in pill form or incorporated as an ingredient in foodstuff 
such as cookies, pretzels or the like. 
Although the invention has been shown in connection with certain specific 
embodiments, it will be readily apparent to those skilled in the art that 
various changes can be made to suit requirements without departing from 
the spirit and scope of the invention.