Therapeutic compositions and methods of use

According to the present invention, there is provided an admixture of sorbic acid, malic acid, fumaric acid, crotonic acid, and optionally aconitic acid. Further contemplated by the present invention are methods for treating various pathological conditions such as viral infections, acidosis, tumors, and bacterial and fungal infections by administering various therapeutically effective amounts of an admixture described above.

FIELD OF THE INVENTION 
The invention relates to admixture compositions of short chain organic 
fatty acids which have biological effects. For example, they are useful in 
the treatment of numerous pathologies that affect mammals. The invention 
also relates to methods of treatment of such pathologies, wherein 
appropriate therapeutically effective amounts of the compositions are 
administered. 
BACKGROUND OF THE INVENTION 
A wide variety of carboxylic acid containing compositions are biologically 
active, and several short chain organic acids which are naturally 
occurring acids have previously been implicated in the treatment of 
various pathological conditions. For example, Nordman, U.S. Pat. No. 
3,291,689, discloses the treatment of hepatic ammonia intoxication with a 
mixture of L-arginine and malic acid. U.S. Pat. No. 3,718,664 discloses 
the use of thioctic acid derivatives in the treatment of acidosis. 
Sloan, U.S. Pat. No. 4,381,307, discloses soft tertiary amine ester 
derivatives that have biological effects, while U.S. Pat. No. 4,760,078 
discloses a 1,2, dithiol-3-thione derivative that has an immunomodulating 
effect. 
Rubenstein, U.S. Pat. No. 4,971,760, discloses the sterilization of blood, 
blood constituents, and transplant tissues with a disinfectant that 
includes lactic acid and sodium chlorite. Naphtholic acid derivatives were 
discovered to be useful in enhancing oxygen availability to mammalian 
tissue by Suh et al., U.S. Pat. No. 5,015,663. 
Hoffiler et al., U.S. Pat. No. 5,043,357, disclose a virucidal agent that 
is predominantly comprised of ethanol and/or alcohol but which includes a 
minor amount of a short chain organic acid. Further, U.S. Pat. No. 
5,093,140 discloses an aqueous bactericide that contains organic acids. 
This solution is used in the scalding or washing stages of meat dressing. 
Kross et al., U.S. Pat. No. 5,100,652 disclose an oral disinfectant that 
contains organic acids. See also Comroe et al., The Lung, 1955 Yearbook of 
Medical Publications Inc., Chptr. 4; John West, Respiratory Physiology, 
The Williams & Wilkins Co., Chptr. 6; Hypoxia, Metabolic Acidosis and 
Circulation, ARTCF . . . Oxford; Arnold et al, "Excessive Intracellular 
Acidosis of Skeletal Muscle on Exercise in a Patient with a Post-Viral 
Exhaustion/Fatigue Syndrome" The Lancet: Jun. 23, 1984; Schweckendiek, W., 
"Heilung von Psoraiasis vulgaris", Med. SschS, 13:103-4, 1959; G. E. 
Abraham et al., "Rationale for the Use of Magnesium and Malic Acid", 
Journal of Nutritional Medicine, 3:49-49 (1992); Kuroda, Z. and Akano, M., 
"Antitumor and Anti-intoxication Activities of Fumaric Acid in Cultured 
Cells", Gann, 727:77-782 (1981); Chemical Abstracts 98945Y, 116, March 
1992 "Antifungal Activity of Fumarates in Mice Infected with C. Albicans"; 
Bauer et al., "Clinishe Wochenschrift 1991 69(2), pp. 722-4; for further 
details of several pathological processes which are susceptible to 
treatment according to the present invention. 
It has now been discovered that admixtures of specific short chain organic 
acids have broad therapeutic effects. 
SUMMARY OF THE INVENTION 
According to the present invention, there is provided a composition which 
comprises an admixture of sorbic acid, aconitic acid, malic acid, fumaric 
acid, crotonic acid, and optionally aconitic acid. 
Further contemplated by the present invention are methods for treating 
various pathological conditions such as viral infections including, but 
not limited to, cytomegalovirus and hepatitis infections; acidosis; 
tumors; and bacterial and fungal infections, by administering various 
therapeutically effective amounts of the admixture described above. 
DETAILED DESCRIPTION OF THE INVENTION 
Organic acids are a large group of compounds, many of which can be derived 
from proteins, carbohydrates, and fats. Although many occur naturally, 
many can be synthetically produced without great difficulty. 
Sorbic acid is a crystalline diolefinic acid having the formula C.sub.6 
H.sub.8 O.sub.2. Typically, it is used as mold and yeast inhibitor; a 
fungistatic agent for foods, especially cheeses; or to improve the 
characteristics of drying oils, the gloss of alkyd type coatings, or the 
milling properties of cold rubber. 
Aconitic acid is a white crystalline acid having the formula C.sub.6 
H.sub.6 O.sub.6. Typically, it is used in manufacturing itaconic acid or 
as a plasticizer for buna rubber and plastics. 
Malic acid is a crystalline dicarboxylic acid having the formula C.sub.4 
H.sub.4 O.sub.5 and is typically found in three isomeric crystalline 
forms. 
Fumaric acid is a crystalline unsaturated dicarboxylic acid having the 
formula C.sub.4 H.sub.4 O.sub.4. Fumaric acid is typically used as a 
substitute for tartaric acid in beverages and baking powders, as a 
replacement or partial replacement of citric acid in soft drinks, and as 
an antioxidant. 
Crotonic acid is an unsaturated aliphatic acid having the formula C.sub.4 
H.sub.6 O.sub.2. It exists in the crystalline form, and is typically used 
in the manufacture of copolymers with vinylacetate that are used in 
lacquers and paper sizing, and in the manufacture of softening agents for 
synthetic rubber. 
It has now been discovered that these acids, in combination, have 
significant therapeutic value. The admixtures of these acids are prepared 
by conventional means known to those skilled in the skilled art. The order 
of addition of the acids to the admixture does not affect the final 
composition. 
Although a wide range of proportions of the individual components of the 
admixture is effective, in a typical composition, each of the four 
required individual acids will independently comprise from about 1 to 
about 50 parts by weight and, optionally, aconitic acid will comprise from 
0 to about 50 parts by weight, based upon 100 parts by weight of the five 
acids combined. Preferably, sorbic acid comprises from about 5 to about 40 
parts by weight, aconitic acid comprises from 0 to about 50 parts by 
weight, malic acid comprises from about 1 to about 30 parts by weight, 
fumaric acid comprises from about 1 to about 30 parts by weight, and 
crotonic acid comprises from about 1 to about 30 parts by weight, based 
upon 100 parts by weight of the five acids combined. Most preferably, 
sorbic acid comprises from about 20 to about 30 parts by weight, aconitic 
acid comprises from about 35 to about 45 parts by weight, malic acid 
comprises from about 10 to about 20 parts by weight, fumaric acid 
comprises from about 10 to about 20 parts by weight, and crotonic acid 
comprises from about 1 to about 20 parts by weight, based upon 100 parts 
by weight of the five acids combined. Special mention is made of an 
admixture wherein sorbic acid comprises about 24 parts by weight, aconitic 
acid comprises about 40 parts by weight, malic acid comprises about 16 
parts by weight, fumaric acid comprises about 16 parts by weight, and 
crotonic acid comprises about 4 parts by weight, based upon 100 parts by 
weight of the five acids combined. 
Suitable pharmaceutically active carriers may be combined with the 
admixture. Such carriers include those known to those skilled in the art 
and can be added at any point in the mixing process. If a pharmaceutically 
active carrier is used, the carrier will comprise from about 1 to about 99 
parts by weight and the acid admixture will comprise from about 99 to 
about 1 part by weight, based upon 100 parts by weight of acid admixture 
and carrier combined. 
The acid admixture with or without the carrier can be formulated into 
dosage unit forms including, but not limited to, capsules and tablets. 
Methods of preparation of dosage unit forms would be known to those 
skilled in the art. Agents which facilitate the manufacture and/or use of 
such dosage unit forms may be added such as plasticizers, lubricants, 
excipients, diluents and the like. 
Methods of treatment of various pathologies contemplated by the present 
invention involve the administration of therapeutically effective amounts 
of the admixture to mammals in need of such treatment. Administration is 
by means known to those skilled in the art. Preferably, administration is 
systemic and most preferably, it is oral. 
Pathologies which are effectively treated with the organic acid admixture 
described herein include but are not limited to acidosis; viral infections 
including, but not limited to, cytomaglovirus and hepatitis infections; 
tumors; and bacterial and fungal infections, including but not limited to 
S. aureus, E. coli, and C. albicans. 
All of these pathologies are well known to those skilled in the art, and 
their diagnosis would be evident to one skilled in the art. For example, 
acidosis is reflected by abnormally acidic blood pH and/or urine pH. 
Normal blood pH ranges from about 7.3 to about 7.38, and normal urine pH 
ranges from about 6 to about 8. Viral, bacterial, and fungal infections 
can be confirmed by appropriate assay methods. Other diseases such as 
chronic fatigue syndrome are only diagnosed through clinical observations. 
Various opportunistic infections which are characteristic of AIDS 
including, but not limited to, cytomegalovirus, hepatitis, and the like 
are effectively treated as described herein. 
The amount of the admixture necessary to treat acidosis is an anti-acidosis 
effective amount. Similarly, the amounts of the admixture necessary to 
treat a viral infection, and particularly hepatitis and cytomegalovirus 
infections are an anti-viral effective amount, and particularly 
anti-hepatitis and an anti-cytomegalovirus effective amounts. The amount 
required to treat tumors, and bacterial and fungal infections are 
anti-tumor, anti-bacterial, and anti-fungal effective amounts, 
respectively. 
The actual amounts of the admixture to be administered will depend 
independently upon the age, weight, sex, sensitivity, medical condition, 
including but not limited to stage of a particular infection or disease, 
or the like, of an individual. However, the amount will be a safe 
non-toxic amount. 
These amounts can be determined by experimentation well-known in the art 
such as by establishing a matrix of dosages and frequencies and assigning 
a group of experimental subjects to each point in the matrix. Typically, 
that amount will range from about 1000 to about 2000 milligrams per day 
for a 150 pound man and preferably about 1500 milligrams per day for a 150 
pound man. Appropriate dosages for different body weights can be 
calculated from this. 
A daily dosage identifies the average amount of the mixture administered to 
an individual. Although the daily dosage may actually be administered 
daily, it need not be administered daily. The daily dosage is merely an 
average dosage that an individual receives when the mixture is 
administered over a period of time. The daily dosage can be administered 
in divided portions so that the total amount administered is the daily 
dosage.

DESCRIPTION OF THE PREFERRED EMBODIMENTS 
The following examples illustrate the invention without limitation. All 
amounts are given as parts by weight (pbw). 
EXAMPLE 1 
An admixture of 24 parts by weight of sorbic acid, 40 parts by weight of 
aconitic acid, 16 parts by weight of malic acid, 16 parts by weight of 
fumaric acid, and 4 parts by weight of crotonic acid was applied, in 
various dilutions, to cut filter paper, and the filter paper was overlayed 
on plates streaked with bacteria. Results are illustrated in Table 1. 
TABLE 1 
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Bacterial Assays 
Admix- Pseudo- 
ture monas Escheri- 
Dilu- Asper- Staph. aeru- chia Candida 
tion gillus aureus ginosa coli albicans pH 
______________________________________ 
Full - 1.6 cm - 1.1 cm 1.6 cm 2.2 
Strength 
1:10 - - - - - 4.6 
1:100 - - - - - 5.2 
______________________________________ 
The (-) negative means that no inhibition was observed. 
EXAMPLE 2 
A patient had a temperature of 101.degree. F. and symptoms of acute urinary 
and prostate infection. The patient was treated with 1500 mg/day of an 
admixture of 24 pbw of sorbic acid, 40 pbw of aconitic acid, 16 pbw of 
malic acid, 16 parts by weight of fumaric acid, and 4 pbw of crotonic acid 
based upon 100 pbw of total admixture for ten days. Symptoms and 
temperature subsided by the 4th or 5th day. Antimicrobial and alkalizing 
activity and therapeutic effects of the admixture are illustrated in Table 
2. Cultures which were positive for streptococci were negative by the 
third week. 
TABLE 2 
______________________________________ 
Urinary Infection 
Initial Treatment 
Day 
Laboratory Test Day 1 10 Day 32 
______________________________________ 
URINALYSIS 
pH 5.5 8 6 
Protein 1+ 0 0 
WBC 20-30 2-4 2-4 
Bacteria many 0 few 
Urine Culture Strept. + .+-. 0 
Blood PSA (normal 0-4) 
21.9 5.1 3.22 
WBC 10.4 6.1 
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EXAMPLE 3 
Six HIV positive individuals with average urine pH at breakfast, lunch, and 
dinner of 6.0, as measured by pHydrion tape, were administered 1500 mg/day 
of an admixture of 24 pbw of sorbic acid, 40 pbw of aconitic acid, 16 pbw 
of malic acid, 16 pbw of fumaric acid, and 4 pbw of crotonic acid. 
After one month, average urine pH was raised to 6.3. Previously, it was 
fixed at 6.0. Results are illustrated in Table 3 below. 
TABLE 3 
______________________________________ 
Acidosis 
Urine pH Urine pH 
Urine pH 
Patient Breakfast Lunch Dinner 
______________________________________ 
"A" 6.0 6.2 6.2 
6.0 6.2 6.0 
6.4 6.2 6.2 
"B" 6.2 6.4 6.2 
6.0 6.0 6.4 
6.2 6.4 6.2 
"C" 6.0 6.0 6.2 
6.0 6.2 6.0 
6.0 6.0 6.2 
"D" 6.0 6.0 6.2 
6.0 6.2 6.2 
6.0 6.0 6.2 
"E" 6.0 6.4 6.4 
6.2 6.2 6.2 
6.0 6.0 6.2 
"F" 6.0 6.4 6.4 
6.2 6.2 6.2 
6.0 6.2 6.2 
______________________________________ 
EXAMPLE 4 
A patient with urine culture positive for cytomegalovirus infection was 
treated with 1500 mg/day of an admixture of 24 pbw of sorbic acid, 40 pbw 
of aconitic acid, 16 pbw of malic acid, 16 pbw of fumaric acid, and 4 pbw 
of crotonic acid, based upon 100 pbw of total admixture for six weeks. 
After six weeks, a cytomegalovirus urine culture was negative. 
EXAMPLE 5 
A patient having cancer of the prostate with a PSA of 17 was treated with 
Zolodex.TM.. PSA was lowered to 2.4. Seven months after Zolodex treatment 
was discontinued, PSA rose to 13. 
The patient then received 1500 mg/day of an admixture of 24 pbw of sorbic 
acid, 40 pbw of aconitic acid, 16 pbw of malic acid, 16 pbw of fumaric 
acid, and 4 pbw of crotonic acid based upon 100 pbw of total admixture, 
and PSA was lowered to 1.65. Nightly urination decreased from 5-6 
times/night to two times/night and the patient's general energy increased 
80%. 
EXAMPLE 6 
A patient was diagnosed with a pancreatic mass. The patient also had 
nausea, pain, an inability to eat, weight loss, and weakness. 
The patient, who weighed about less than eighty pounds, was given 50 mg/day 
of an admixture of 24 pbw of sorbic acid, 40 pbw of aconitic acid, 16 pbw 
of malic acid, 16 pbw of fumaric acid, and 4 pbw of crotonic acid based 
upon 100 pbw of total admixture for two months. The patient's nausea 
abated, her appetite improved, she slept more restfully, and she did not 
urinate as frequently at night. The epigastric mass was no longer 
palpable. 
EXAMPLE 7 
A patient underwent a resection of a recurrent bladder tumor, at which 
time, the pathological specimen revealed Grade II/III moderately 
differentiated papillary transitional cell carcinoma with invasion of the 
muscularis of the urinary bladder and of the prostate. The patient was 
placed on a protocol of 1500 mg/day of an admixture of 24 pbw of sorbic 
acid, 40 pbw of aconitic acid, 16 pbw of malic acid, 16 pbw of fumaric 
acid, and 4 pbw of crotonic acid based upon 100 pbw of total admixture. A 
repeat cystoscopy two months later showed only several small papillary 
lesions in the urinary bladder and a small lesion in the prostatic 
urethra. Rectal examination was unremarkable. CAT SCAN confirmed the 
improvement observed by cystoscopy. 
EXAMPLE 8 
A patient was diagnosed with long standing Hepatitis C. The patient was 
treated with 1500 mg/day of an admixture of 24 pbw of sorbic acid, 40 pbw 
of aconitic acid, 16 pbw of malic acid, 16 pbw of fumaric acid, and 4 pbw 
of crotonic acid based upon 100 pbw of total admixture. After one month, 
pain in the patient's side subsided. The patient was no longer nauseated 
and generally felt better. His SGOT laboratory value went down from 68 to 
37. 
EXAMPLE 9 
An admixture of 24 pbw of sorbic acid, 40 pbw of aconitic acid, 16 pbw of 
malic acid, 16 pbw of fumaric acid, and 4 pbw of crotonic acid based upon 
100 pbw of total admixture was tested for virucidal effectiveness against 
Cytomegalovirus. The viral inoculum was diluted 1:10 with the admixture. 
The test material/virus mixture was sampled at 3 and 6 minutes. The 
treated viral mixture was neutralized and titered. An initial viral titer 
was determined. 
The challenge organism was Cytomegalovirus, (CMV) and the host cell line 
was MRC-5, ATCC CCL-171. Reagents and media were EMEM with Earle's salts 
supplemented with heat inactivated fetal bovine serum (FBS), glutamine, 
and penicillin streptomycin (MEM complete); sterile phosphate buffered 
saline; and newborn calf serum (NBCS). 
The virus stock culture was titered and adjusted to contain approximately 
10.sup.7 CCID.sub.50 /ml (50% cell-culture infectious doses per 
milliliter) and stored in liquid nitrogen until use. The virus stock was 
thawed and was diluted 10-fold in MEM through 10.sup.7. One ml aliquots of 
appropriate dilutions were plated onto 5 confluent MRC-5 monolayers. The 
cells were incubated at 37.+-.1.degree. C., in 5% CO.sub.2, for 2 hours to 
allow the virus to adsorb to the cells. After adsorption, the medium was 
withdrawn from the monolayers, the cells were washed once with PBS, and 
refed with MEM. The monolayers were incubated for 14 days at 
37.+-.1.degree. C., in 5% CO.sub.2, and subsequently were observed for 
virus-specific cytopathogenic effects (CPE). 
The test solution was tested by combining the virus with the admixture at a 
1:10 dilution. This suspension was agitated continuously at a constant 
temperature of 25.degree. C. and samples were withdrawn at 3 and 6 minutes 
to determine a survivor curve and to calculate a D-value. 
The sampled suspensions were neutralized with a 1:1 dilution in new-born 
calf serum and were diluted ten-fold in MEM. One ml aliquots of the 
neutralized mixture were plated onto MRC-5 monolayers (four wells per 
dilution) and were incubated at 37.-+.1.degree. C. in a 5% CO.sub.2 in air 
atmosphere for 2 hours. Following the incubation period, the fluids were 
removed. The monolayers were washed with PBS, refed MEM and were incubated 
at 37.+-.1.degree. C. in a 5% CO.sub.2 in air atmosphere for thirteen 
days. 
Control virus stock was titered at the time of the test as described, to 
confirm that the titer was in an acceptable range. The maintenance of the 
virus titer was evaluated using PBS instead of the test solution. Samples 
were taken less than 15 seconds after the initiation of the test (zero 
time control) and after 6 minutes (final time control). The average 
CCID.sub.50 /ml for each titer was determined by the Reed and Muench 
method. American Journal of Hygiene, 1938, 27:493. The D-value was 
calculated by the Stumbo method. Stumbo, C. R., Thermobacteriology in food 
processing, New York Academic Press (1973), pp. 235-247. 
No CPE was observed at 3 minutes. A D-value calculated using this time 
resulted in a D-value of 45 seconds. This is an estimate since the first 
sampling was done at 3 minutes (no data were collected at less than 3 
minutes). 
When a survivor curve was determined in this test, the admixture provided 
data used to calculate a D-value of 45 seconds (estimated). A conclusion 
can be drawn that the admixture was antiviral against cytomegalovirus. 
Results are illustrated in Tables 4, 5, and 6 below. 
TABLE 4 
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Test and Control Titers Expressed as CPE 
Day 13 Results 
TEST 
PBS SOLUTION 
DILUTION 
VIRUS STOCK Zero 6 min 3 min 
6 min 
______________________________________ 
10.sup.-1 
NA ++++ ++++ ---- ---- 
10.sup.-2 ++++ ++++ ++++ ---- ---- 
10.sup.-3 ++++ ++++ ++++ ---- ---- 
10.sup.-4 ++++ +-+- +-+- ---- ---- 
10.sup.-5 ++++ ---- ---- ---- ---- 
10.sup.-6 ---- ---- ---- ---- ---- 
10.sup.-7 ---- N/A N/A N/A N/A 
CCID.sub.50 /ml 3.2 .times. 10.sup.5 1 .times. 10.sup.4 1 .times. 
10.sup.4 
______________________________________ 
+ = CPE observed 
- = No CPE observed 
CT = Cytotoxicity observed 
O = No Cytotoxicity observed 
TABLE 5 
______________________________________ 
Control and Test Titers at Time Intervals 
After Exposure to Scam Disinfectant 
SAMPLE CCID.sub.50 /ml 
______________________________________ 
Sample 3.2 .times. 10.sup.5 
PBS at Zero time 1 .times. 10.sup.4 
PBS at 6 minutes 1 .times. 10.sup.4 
3 minutes No CPE observed 
6 minutes No CPE observed 
______________________________________ 
TABLE 6 
______________________________________ 
Determination of the D-Value 
______________________________________ 
#STR1## 
______________________________________ 
Tx = end of time interval 
To = beginning of time interval 
AO = CCID.sub.50 /ml at To 
AX = CCID.sub.50 /ml at Tx 
EXAMPLE 10 
A 36 year old male with well controlled HIV infection had progressively 
deteriorating kidney function from no demonstrable cause. He averaged 
5.times. nocturia and ran a consistently 3 plus albuminuria with urinary 
specific gravity fixed at 1.005 (normal to 1.030). 
After approximately 4 months of 1500 mg/day of admixture of 24 pbw of 
sorbic acid, 40 pbw of aconitic acid, 16 pbw of malic acid, 16 pbw of 
fumaric acid, and 4 pbw of crotonic acid based upon 100 pbw of total 
admixture, his kidney function normalized. His three plus albuminuria 
gradually became negative; he became able to sleep through nights, and the 
specific gravity of his urine by concentration test increased to 1.0020. 
His urinary pH, which had been fixed at 5-6 resumed a normal pattern of 
morning aciduria, increasing to neutral or alkaline during the day 
according to his diet and activity. 
Accompanying this was a total regression of his long-existing bilateral 
massively swollen parotid glands; the non-specific diagnosis derived from 
multiple biopsies had been cystic parotitis. The presumptive 
retro-diagnosis for parotitis and the kidney insufficiency was severe 
cytomegalovirus (CMV) infection, suggested by high titers of CMV 
antibodies. 
The above mentioned patents, test methods, and publications are hereby 
incorporated by reference in their entirety. 
Many variations of the present invention will suggest themselves to those 
skilled in the art in light of the above detailed description. All such 
obvious variations are within the full intended scope of the appended 
claims.